{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:49:42Z","timestamp":1787028582951,"version":"build-2736575974"},"reference-count":233,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2026,7,4]],"date-time":"2026-07-04T00:00:00Z","timestamp":1783123200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Agriculture and Rural Development","award":["DHR.hn.070.1.2026"],"award-info":[{"award-number":["DHR.hn.070.1.2026"]}]},{"award":["DHR.hn.070.1.2026"],"award-info":[{"award-number":["DHR.hn.070.1.2026"]}],"id":[{"id":"https:\/\/ror.org\/05rdz5425","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Genome editing, particularly CRISPR\/Cas-based systems, has emerged as a key tool for functional genomics and trait improvement in flax (Linum usitatissimum L.), an important fiber and oilseed crop. This review focuses specifically on flax as an emerging target species and distinguishes experimentally validated applications from approaches adapted from model plants. Recent progress includes the characterization of endogenous U6 promoters, which improved guide RNA expression and contributed to enhanced genome editing performance under optimized conditions. Reported studies demonstrate efficient targeted mutagenesis in flax; however, editing outcomes remain strongly dependent on genotype, construct design, and regeneration capacity, and stable homozygous edited lines are still limited. Target genes include pathways involved in lignin and cellulose biosynthesis, fatty acid metabolism, and stress responses, influencing fiber quality, oil composition, and stress adaptation. Despite current bottlenecks such as low homologous recombination efficiency and regeneration constraints, base editing, prime editing, and multiplex CRISPR systems provide promising avenues for precision breeding in flax.<\/jats:p>","DOI":"10.3390\/ijms27136012","type":"journal-article","created":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T13:43:14Z","timestamp":1783345394000},"page":"6012","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Genome Editing Approaches in Flax (Linum usitatissimum L.): From Tools to Trait Improvement"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9720-7881","authenticated-orcid":false,"given":"Marta","family":"Podralska","sequence":"first","affiliation":[{"name":"Institute of Natural Fibres and Medicinal Plants\u2014National Research Institute, Wojska Polskiego 71B, 60-630 Pozna\u0144, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aleksandra","family":"G\u00f3rska","sequence":"additional","affiliation":[{"name":"Institute of Natural Fibres and Medicinal Plants\u2014National Research Institute, Wojska Polskiego 71B, 60-630 Pozna\u0144, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariusz","family":"Kaczmarek","sequence":"additional","affiliation":[{"name":"Institute of Natural Fibres and Medicinal Plants\u2014National Research Institute, Wojska Polskiego 71B, 60-630 Pozna\u0144, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"44304","DOI":"10.1038\/srep44304","article-title":"Targeted Mutagenesis in Cotton (Gossypium hirsutum L.) Using the CRISPR\/Cas9 System","volume":"7","author":"Chen","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1126\/science.1225829","article-title":"A Programmable Dual-RNA\u2013Guided DNA Endonuclease in Adaptive Bacterial Immunity","volume":"337","author":"Jinek","year":"2012","journal-title":"Science"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1146\/annurev-arplant-042811-105552","article-title":"Plant Genome Engineering with Sequence-Specific Nucleases","volume":"64","author":"Voytas","year":"2013","journal-title":"Annu. Rev. Plant Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1509","DOI":"10.1126\/science.1178811","article-title":"Breaking the Code of DNA Binding Specificity of TAL-Type III Effectors","volume":"326","author":"Boch","year":"2009","journal-title":"Science"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.biotechadv.2014.12.006","article-title":"The CRISPR\/Cas9 System for Plant Genome Editing and Beyond","volume":"33","author":"Bortesi","year":"2015","journal-title":"Biotechnol. Adv."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1146\/annurev-arplant-050718-100049","article-title":"CRISPR\/Cas Genome Editing and Precision Plant Breeding in Agriculture","volume":"70","author":"Chen","year":"2019","journal-title":"Annu. Rev. Plant Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1126\/science.1183700","article-title":"Breeding Technologies to Increase Crop Production in a Changing World","volume":"327","author":"Tester","year":"2010","journal-title":"Science"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"682","DOI":"10.1111\/nph.14702","article-title":"Towards CRISPR\/Cas Crops\u2014Bringing Together Genomics and Genome Editing","volume":"216","author":"Scheben","year":"2017","journal-title":"New Phytol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Khatodia, S., Bhatotia, K., Passricha, N., Khurana, S.M.P., and Tuteja, N. (2016). The CRISPR\/Cas Genome-Editing Tool: Application in Improvement of Crops. Front. Plant Sci., 7.","DOI":"10.3389\/fpls.2016.00506"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1621","DOI":"10.1016\/j.cell.2021.01.005","article-title":"Genome Engineering for Crop Improvement and Future Agriculture","volume":"184","author":"Gao","year":"2021","journal-title":"Cell"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"102191","DOI":"10.1016\/j.pmpp.2023.102191","article-title":"CRISPR\/Cas9-Based Genome Editing: A Revolutionary Approach for Crop Improvement and Global Food Security","volume":"129","author":"Ali","year":"2024","journal-title":"Physiol. Mol. Plant Pathol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1038\/nbt.2199","article-title":"High-Efficiency TALEN-Based Gene Editing Produces Disease-Resistant Rice","volume":"30","author":"Li","year":"2012","journal-title":"Nat. Biotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.jare.2024.08.024","article-title":"CRISPR\/Cas Genome Editing in Soybean: Challenges and New Insights to Overcome Existing Bottlenecks","volume":"73","author":"Arraes","year":"2025","journal-title":"J. Adv. Res."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Tiwari, J.K., Singh, A.K., and Behera, T.K. (2023). CRISPR\/Cas Genome Editing in Tomato Improvement: Advances and Applications. Front. Plant Sci., 14.","DOI":"10.3389\/fpls.2023.1121209"},{"key":"ref_15","first-page":"4304","article-title":"Flax (Linum usitatissimum L.): Current Uses and Future Applications","volume":"4","author":"Jhala","year":"2010","journal-title":"Aust. J. Basic Appl. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1633","DOI":"10.1007\/s13197-013-1247-9","article-title":"Flax and Flaxseed Oil: An Ancient Medicine & Modern Functional Food","volume":"51","author":"Goyal","year":"2014","journal-title":"J. Food Sci. Technol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Gao, Z., Cao, Q., and Deng, Z. (2024). Unveiling the Power of Flax Lignans: From Plant Biosynthesis to Human Health Benefits. Nutrients, 16.","DOI":"10.3390\/nu16203520"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Nowak, W., and Jeziorek, M. (2023). The Role of Flaxseed in Improving Human Health. Healthcare, 11.","DOI":"10.3390\/healthcare11030395"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e114","DOI":"10.1002\/efd2.114","article-title":"A Review on Flaxseeds: Nutritional Profile, Health Benefits, Value Added Products, and Toxicity","volume":"4","author":"Pramanik","year":"2023","journal-title":"eFood"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1111\/j.1365-313X.2012.05093.x","article-title":"The Genome of Flax (Linum usitatissimum) Assembled de Novo from Short Shotgun Sequence Reads","volume":"72","author":"Wang","year":"2012","journal-title":"Plant J."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Li, Z., Chi, H., Liu, C., Zhang, T., Han, L., Li, L., Pei, X., and Long, Y. (2021). Genome-Wide Identification and Functional Characterization of LEA Genes during Seed Development Process in Linseed Flax (Linum usitatissimum L.). BMC Plant Biol., 21.","DOI":"10.1186\/s12870-021-02972-0"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1089\/crispr.2024.0064","article-title":"Establishment of a CRISPR-Cas9-Mediated Genome Editing System in Flax","volume":"8","author":"Wang","year":"2025","journal-title":"CRISPR J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"S237","DOI":"10.1134\/S102279542314003X","article-title":"Improving Genome Assembly of Flax Line 3896 with High-Precision Illumina Reads","volume":"59","author":"Dvorianinova","year":"2023","journal-title":"Russ. J. Genet."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1038\/nrg2842","article-title":"Genome Editing with Engineered Zinc Finger Nucleases","volume":"11","author":"Urnov","year":"2010","journal-title":"Nat. Rev. Genet."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1843","DOI":"10.1126\/science.1204094","article-title":"TAL Effectors: Customizable Proteins for DNA Targeting","volume":"333","author":"Bogdanove","year":"2011","journal-title":"Science"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Raina, A., Sahu, P.K., Laskar, R.A., Rajora, N., Sao, R., Khan, S., and Ganai, R.A. (2021). Mechanisms of Genome Maintenance in Plants: Playing It Safe with Breaks and Bumps. Front. Genet., 12.","DOI":"10.3389\/fgene.2021.675686"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1111\/tpj.12338","article-title":"Synthetic Nucleases for Genome Engineering in Plants: Prospects for a Bright Future","volume":"78","author":"Puchta","year":"2014","journal-title":"Plant J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1007\/s42994-019-00009-7","article-title":"Precise Gene Replacement in Plants through CRISPR\/Cas Genome Editing Technology: Current Status and Future Perspectives","volume":"1","author":"Li","year":"2020","journal-title":"aBIOTECH"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e11314","DOI":"10.1002\/aps3.11314","article-title":"Considerations in Adapting CRISPR\/Cas9 in Nongenetic Model Plant Systems","volume":"8","author":"Shan","year":"2020","journal-title":"Appl. Plant Sci."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Rozov, S.M., Permyakova, N.V., and Deineko, E.V. (2019). The Problem of the Low Rates of CRISPR\/Cas9-Mediated Knock-Ins in Plants: Approaches and Solutions. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20133371"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Naeem, M., and Alkhnbashi, O.S. (2023). Current Bioinformatics Tools to Optimize CRISPR\/Cas9 Experiments to Reduce Off-Target Effects. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24076261"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1186\/s43141-020-00036-8","article-title":"The Present and Potential Future Methods for Delivering CRISPR\/Cas9 Components in Plants","volume":"18","author":"Sandhya","year":"2020","journal-title":"J. Genet. Eng. Biotechnol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e202400457","DOI":"10.1002\/biot.202400457","article-title":"Advancements in Plant Gene Editing Technology: From Construct Design to Enhanced Transformation Efficiency","volume":"19","author":"Yuan","year":"2024","journal-title":"Biotechnol. J."},{"key":"ref_34","first-page":"1510","article-title":"Advancing Crop Transformation in the Era of Genome Editing","volume":"28","author":"Altpeter","year":"2016","journal-title":"Plant Cell"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Han, X., Deng, Z., Liu, H., and Ji, X. (2025). Current Advancement and Future Prospects in Simplified Transformation-Based Plant Genome Editing. Plants, 14.","DOI":"10.3390\/plants14060889"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1998","DOI":"10.1105\/tpc.16.00124","article-title":"Morphogenic Regulators Baby Boom and Wuschel Improve Monocot Transformation","volume":"28","author":"Lowe","year":"2016","journal-title":"Plant Cell"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1274","DOI":"10.1038\/s41587-020-0703-0","article-title":"A GRF\u2013GIF Chimeric Protein Improves the Regeneration Efficiency of Transgenic Plants","volume":"38","author":"Debernardi","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1038\/s41587-019-0337-2","article-title":"Plant Gene Editing through de Novo Induction of Meristems","volume":"38","author":"Maher","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1038\/s41586-018-0785-8","article-title":"A Male-Expressed Rice Embryogenic Trigger Redirected for Asexual Propagation through Seeds","volume":"565","author":"Khanday","year":"2019","journal-title":"Nature"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1038\/s41477-026-02237-4","article-title":"High-Efficiency, Transgene-Free Plant Genome Editing by Viral Delivery of an Engineered TnpB","volume":"12","author":"Nagalakshmi","year":"2026","journal-title":"Nat. Plants"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"692","DOI":"10.1038\/s41586-021-04058-1","article-title":"Transposon-Associated TnpB Is a Programmable RNA-Guided DNA Endonuclease","volume":"599","author":"Karvelis","year":"2021","journal-title":"Nature"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1126\/science.abj6856","article-title":"The Widespread IS200\/IS605 Transposon Family Encodes Diverse Programmable RNA-Guided Endonucleases","volume":"374","author":"Kannan","year":"2021","journal-title":"Science"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"620","DOI":"10.1038\/s41477-020-0670-y","article-title":"Multiplexed Heritable Gene Editing Using RNA Viruses and Mobile Single Guide RNAs","volume":"6","author":"Ellison","year":"2020","journal-title":"Nat. Plants"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1787","DOI":"10.1016\/j.molp.2021.07.010","article-title":"Highly Efficient Heritable Genome Editing in Wheat Using an RNA Virus and Bypassing Tissue Culture","volume":"14","author":"Li","year":"2021","journal-title":"Mol. Plant"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2954","DOI":"10.1038\/s41596-019-0208-9","article-title":"Carbon Nanotube\u2013Mediated DNA Delivery without Transgene Integration in Intact Plants","volume":"14","author":"Demirer","year":"2019","journal-title":"Nat. Protoc."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Shang, Y., Hasan, M.K., Ahammed, G.J., Li, M., Yin, H., and Zhou, J. (2019). Applications of Nanotechnology in Plant Growth and Crop Protection: A Review. Molecules, 24.","DOI":"10.3390\/molecules24142558"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"956","DOI":"10.1038\/s41477-017-0063-z","article-title":"Pollen Magnetofection for Genetic Modification with Magnetic Nanoparticles as Gene Carriers","volume":"3","author":"Zhao","year":"2017","journal-title":"Nat. Plants"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"10966","DOI":"10.1038\/s41598-018-29298-6","article-title":"Library Screening of Cell-Penetrating Peptide for BY-2 Cells, Leaves of Arabidopsis, Tobacco, Tomato, Poplar, and Rice Callus","volume":"8","author":"Numata","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"13274","DOI":"10.1038\/ncomms13274","article-title":"Genome Editing in Maize Directed by CRISPR\u2013Cas9 Ribonucleoprotein Complexes","volume":"7","author":"Svitashev","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2034","DOI":"10.1111\/pbi.70028","article-title":"Plant Genetic Transformation: Achievements, Current Status and Future Prospects","volume":"23","author":"Wang","year":"2025","journal-title":"Plant Biotechnol. J."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1196","DOI":"10.1105\/tpc.16.00922","article-title":"A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants","volume":"29","author":"Curtin","year":"2017","journal-title":"Plant Cell"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Xing, H.-L., Dong, L., Wang, Z.-P., Zhang, H.-Y., Han, C.-Y., Liu, B., Wang, X.-C., and Chen, Q.-J. (2014). A CRISPR\/Cas9 Toolkit for Multiplex Genome Editing in Plants. BMC Plant Biol., 14.","DOI":"10.1186\/s12870-014-0327-y"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1836","DOI":"10.1007\/s12033-023-00826-8","article-title":"Agrobacterium-Mediated Transformation for the Development of Transgenic Crops; Present and Future Prospects","volume":"66","author":"Rahman","year":"2024","journal-title":"Mol. Biotechnol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"697","DOI":"10.18699\/VJGB-22-84","article-title":"Agrobacterium-Mediated Transformation of Nicotiana glauca and Nicotiana sylvestris","volume":"26","author":"Khafizova","year":"2022","journal-title":"Vavilov J. Genet. Breed."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"e620","DOI":"10.1002\/cpz1.620","article-title":"Agrobacterium-mediated Genetic Transformation of Cassava","volume":"2","author":"Segatto","year":"2022","journal-title":"Curr. Protoc."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"e58","DOI":"10.1002\/cpz1.58","article-title":"An Efficient Agrobacterium-Mediated Transformation Protocol for Hexaploid and Tetraploid Wheat","volume":"1","author":"Hayta","year":"2021","journal-title":"Curr. Protoc."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Li, Y., Tang, D., Liu, Z., Chen, J., Cheng, B., Kumar, R., Yer, H., and Li, Y. (2022). An Improved Procedure for Agrobacterium-Mediated Transformation of \u2018Carrizo\u2019 Citrange. Plants, 11.","DOI":"10.3390\/plants11111457"},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Shao, M., McCue, K.F., and Thomson, J.G. (2024). Agrobacterium-Mediated Transformation of the Dwarf Soybean MiniMax. Plants, 13.","DOI":"10.3390\/plants13071013"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1007\/s00425-021-03666-5","article-title":"Improved Cotton Transformation Protocol Mediated by Agrobacterium and Biolistic Combined-Methods","volume":"254","author":"Ribeiro","year":"2021","journal-title":"Planta"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1007\/s11248-023-00355-9","article-title":"Development of Agrobacterium-Mediated in Planta Transformation Protocol through Coleoptile in Rice","volume":"32","author":"Ho","year":"2023","journal-title":"Transgenic Res."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1007\/s11248-025-00468-3","article-title":"The Agrobacterium-Mediated Genetic Transformation: A Gateway for Efficient CRISPR\/Cas9 Gene Editing in Leguminous","volume":"34","author":"Sarroukh","year":"2025","journal-title":"Transgenic Res."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"1563","DOI":"10.1007\/s12033-023-00788-x","article-title":"Agrobacterium Tumefaciens-Mediated Plant Transformation: A Review","volume":"66","author":"Pouresmaeil","year":"2024","journal-title":"Mol. Biotechnol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"109652","DOI":"10.1016\/j.plaphy.2025.109652","article-title":"Gibberellin Biosynthesis Gene ScGA20 Oxidase Enhances Sugarcane Growth by Modulating Genes Associated with Phytohormone and Growth Processes","volume":"221","author":"Wang","year":"2025","journal-title":"Plant Physiol. Biochem."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"7695","DOI":"10.1038\/s41598-021-86549-9","article-title":"An Improved Biolistic Delivery and Analysis Method for Evaluation of DNA and CRISPR-Cas Delivery Efficacy in Plant Tissue","volume":"11","author":"Miller","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"4760","DOI":"10.1093\/jxb\/erae243","article-title":"Biolistics-Mediated Transformation of Hornworts and Its Application to Study Pyrenoid Protein Localization","volume":"75","author":"Lafferty","year":"2024","journal-title":"J. Exp. Bot."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1007\/s00299-024-03359-9","article-title":"Advancements in Plant Transformation: From Traditional Methods to Cutting-Edge Techniques and Emerging Model Species","volume":"43","author":"Levengood","year":"2024","journal-title":"Plant Cell Rep."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"154122","DOI":"10.1016\/j.jplph.2023.154122","article-title":"Preparation of Leaf Protoplasts from Populus (Populus \u00d7 Xiaohei T.S. Hwang et Liang) and Establishment of Transient Expression System","volume":"291","author":"Yang","year":"2023","journal-title":"J. Plant Physiol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s13007-023-00983-6","article-title":"Establishment of an Efficient Cotton Root Protoplast Isolation Protocol Suitable for Single-Cell RNA Sequencing and Transient Gene Expression Analysis","volume":"19","author":"Zhang","year":"2023","journal-title":"Plant Methods"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1186\/s13007-024-01180-9","article-title":"Optimization of Preparation Conditions for Salsola Laricifolia Protoplasts Using Response Surface Methodology and Artificial Neural Network Modeling","volume":"20","author":"Guo","year":"2024","journal-title":"Plant Methods"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1186\/s13007-025-01471-9","article-title":"PVP-40 Mediated Enhancement of Mesophyll Protoplast Yield and Viability for Transient Gene Expression in Black Huckleberry","volume":"21","author":"Majumder","year":"2025","journal-title":"Plant Methods"},{"key":"ref_71","doi-asserted-by":"crossref","unstructured":"Figueroa-Varela, P., Susunaga-G\u00f3mez, D., Restrepo-Osorio, C., Harms, C., and Villanueva-Mej\u00eda, D. (2023). An Efficient Method for Protoplast-Mediated Production of Transformed Castor Bean (Ricinus communis) Lines. BMC Res. Notes, 16.","DOI":"10.1186\/s13104-023-06414-y"},{"key":"ref_72","doi-asserted-by":"crossref","unstructured":"Hu, Y., Wu, Y., Song, J., Ma, M., Xiao, Y., and Zeng, B. (2024). Advancing Cordyceps Militaris Industry: Gene Manipulation and Sustainable Biotechnological Strategies. Bioengineering, 11.","DOI":"10.3390\/bioengineering11080783"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"e3782","DOI":"10.21769\/BioProtoc.3782","article-title":"Cellulase and Macerozyme-PEG-Mediated Transformation of Moss Protoplasts","volume":"10","author":"Batth","year":"2020","journal-title":"Bio. Protoc."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1038\/s41586-025-08706-8","article-title":"Perception of Viral Infections and Initiation of Antiviral Defence in Rice","volume":"641","author":"Huang","year":"2025","journal-title":"Nature"},{"key":"ref_75","doi-asserted-by":"crossref","unstructured":"Huang, F.-F., Xie, Y.-H., Hu, Q., Huang, M.-X., Zhang, L.-Z., Liu, Z.-H., Li, J., Huang, J.-A., and Xiong, L.-G. (2026). PEG-Mediated Transformation: A Tool for Random Integration or Targeted Gene Replacement in Colletotrichum Camelliae. Front. Microbiol., 17.","DOI":"10.3389\/fmicb.2026.1814272"},{"key":"ref_76","doi-asserted-by":"crossref","unstructured":"Wang, L., Liu, H., Liu, P., Wu, G., Shen, W., Cui, H., and Dai, Z. (2023). Cotyledon Peeling Method for Passion Fruit Protoplasts: A Versatile Cell System for Transient Gene Expression in Passion Fruit (Passiflora edulis). Front. Plant Sci., 14.","DOI":"10.3389\/fpls.2023.1236838"},{"key":"ref_77","doi-asserted-by":"crossref","unstructured":"Subburaj, S., and Agapito-Tenfen, S.Z. (2023). Establishment of Targeted Mutagenesis in Soybean Protoplasts Using CRISPR\/Cas9 RNP Delivery via Electro\u2212transfection. Front. Plant Sci., 14.","DOI":"10.3389\/fpls.2023.1255819"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"106912","DOI":"10.1016\/j.mimet.2024.106912","article-title":"Optimisation of DNA Electroporation Protocols for Different Plant-Associated Bacteria","volume":"220","author":"Kim","year":"2024","journal-title":"J. Microbiol. Methods"},{"key":"ref_79","doi-asserted-by":"crossref","unstructured":"Zhan, C., Hu, J., and Zhang, Y. (2026). Virus-Mediated Genome Editing in Grasses. Trends Biotechnol.","DOI":"10.1016\/j.tibtech.2026.02.019"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"1252","DOI":"10.1038\/s41477-025-02023-8","article-title":"Transgene- and Tissue Culture-Free Heritable Genome Editing Using RNA Virus-Based Delivery in Wheat","volume":"11","author":"Qiao","year":"2025","journal-title":"Nat. Plants"},{"key":"ref_81","doi-asserted-by":"crossref","unstructured":"Zhang, C., Liu, S., Li, X., Zhang, R., and Li, J. (2022). Virus-Induced Gene Editing and Its Applications in Plants. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms231810202"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1007\/s42994-024-00147-7","article-title":"Exploiting Viral Vectors to Deliver Genome Editing Reagents in Plants","volume":"5","author":"Shen","year":"2024","journal-title":"aBIOTECH"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"109922","DOI":"10.1016\/j.virol.2023.109922","article-title":"Considerations in Engineering Viral Vectors for Genome Editing in Plants","volume":"589","author":"Wu","year":"2024","journal-title":"Virology"},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"101992","DOI":"10.1016\/j.pbi.2020.101992","article-title":"Virus-Induced Plant Genome Editing","volume":"60","author":"Oh","year":"2021","journal-title":"Curr. Opin. Plant Biol."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1186\/s13059-015-0796-9","article-title":"High-Frequency, Precise Modification of the Tomato Genome","volume":"16","author":"Baltes","year":"2015","journal-title":"Genome Biol."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"1251","DOI":"10.1111\/tpj.13446","article-title":"High-efficiency Gene Targeting in Hexaploid Wheat Using DNA replicons and CRISPR\/Cas9","volume":"89","author":"Wang","year":"2017","journal-title":"Plant J."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1105\/tpc.113.119792","article-title":"DNA Replicons for Plant Genome Engineering","volume":"26","author":"Baltes","year":"2014","journal-title":"Plant Cell"},{"key":"ref_88","doi-asserted-by":"crossref","unstructured":"Borphukan, B., Khatun, M., Fartyal, D., James, D., and Reddy, M.K. (2025). A Gemini Virus-Derived Autonomously Replicating System for HDR-Mediated Genome Editing of the EPSP Synthase Gene in Indica Rice. Plants, 14.","DOI":"10.3390\/plants14030477"},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1038\/s41565-019-0375-4","article-title":"Chloroplast-Selective Gene Delivery and Expression in Planta Using Chitosan-Complexed Single-Walled Carbon Nanotube Carriers","volume":"14","author":"Kwak","year":"2019","journal-title":"Nat. Nanotechnol."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"e70227","DOI":"10.1111\/ppl.70227","article-title":"Nanoparticles as Tools for Enhancing Plant Resistance to Biotic Stress in the Context of Climate Change","volume":"177","author":"Ruffatto","year":"2025","journal-title":"Physiol. Plant."},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"882","DOI":"10.1016\/j.tibtech.2018.03.009","article-title":"Nanoparticle-Mediated Delivery towards Advancing Plant Genetic Engineering","volume":"36","author":"Cunningham","year":"2018","journal-title":"Trends Biotechnol."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"7079","DOI":"10.1007\/s11033-022-07442-w","article-title":"CRISPR\/Cas9 a Simple, Inexpensive and Effective Technique for Gene Editing","volume":"49","author":"Ferreira","year":"2022","journal-title":"Mol. Biol. Rep."},{"key":"ref_93","doi-asserted-by":"crossref","unstructured":"Veillet, F., Perrot, L., Guyon-Debast, A., Kermarrec, M.-P., Chauvin, L., Chauvin, J.-E., Gallois, J.-L., Mazier, M., and Nogu\u00e9, F. (2020). Expanding the CRISPR Toolbox in P. Patens Using SpCas9-NG Variant and Application for Gene and Base Editing in Solanaceae Crops. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21031024"},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1038\/s41477-019-0375-2","article-title":"Targeted DNA Methylation Represses Two Enhancers of FLOWERING LOCUS T in Arabidopsis Thaliana","volume":"5","author":"Zicola","year":"2019","journal-title":"Nat. Plants"},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"1587","DOI":"10.1016\/j.molp.2019.09.007","article-title":"A PIF7-CONSTANS-Centered Molecular Regulatory Network Underlying Shade-Accelerated Flowering","volume":"12","author":"Zhang","year":"2019","journal-title":"Mol. Plant"},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1080\/21645698.2015.1134405","article-title":"Future of Breeding by Genome Editing Is in the Hands of Regulators","volume":"6","author":"Jones","year":"2015","journal-title":"GM Crops Food"},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"3320","DOI":"10.1073\/pnas.1720692115","article-title":"Regional Pest Suppression Associated with Widespread Bt Maize Adoption Benefits Vegetable Growers","volume":"115","author":"Dively","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1186\/s42397-024-00192-z","article-title":"Emerging Technological Developments to Address Pest Resistance in Bt Cotton","volume":"7","author":"Nagaraj","year":"2024","journal-title":"J. Cotton Res."},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1002\/ps.1008","article-title":"Glyphosate-resistant Crops: History, Status and Future","volume":"61","author":"Dill","year":"2005","journal-title":"Pest. Manag. Sci."},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1186\/s13059-024-03448-8","article-title":"Systemic Evaluation of Various CRISPR\/Cas13 Orthologs for Knockdown of Targeted Transcripts in Plants","volume":"25","author":"Yu","year":"2024","journal-title":"Genome Biol."},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1007\/s42994-025-00228-1","article-title":"Molecular Mechanisms and Crop Improvement Potential of RNA N6-Methyladenosine in Plants","volume":"6","author":"Fu","year":"2025","journal-title":"aBIOTECH"},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1104\/pp.125.3.1157","article-title":"Golden Rice and Beyond","volume":"125","author":"Potrykus","year":"2001","journal-title":"Plant Physiol."},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"506S","DOI":"10.1093\/jn\/132.3.506S","article-title":"Golden Rice: Introducing the \u03b2-Carotene Biosynthesis Pathway into Rice Endosperm by Genetic Engineering to Defeat Vitamin A Deficiency","volume":"132","author":"Beyer","year":"2002","journal-title":"J. Nutr."},{"key":"ref_104","doi-asserted-by":"crossref","unstructured":"Wairich, A., Ricachenevsky, F.K., and Lee, S. (2022). A Tale of Two Metals: Biofortification of Rice Grains with Iron and Zinc. Front. Plant Sci., 13.","DOI":"10.3389\/fpls.2022.944624"},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1016\/j.molp.2019.03.011","article-title":"Improving Plant Genome Editing with High-Fidelity XCas9 and Non-Canonical PAM-Targeting Cas9-NG","volume":"12","author":"Zhong","year":"2019","journal-title":"Mol. Plant"},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1111\/jipb.12886","article-title":"Expanding the Scope of CRISPR\/Cas9-mediated Genome Editing in Plants Using an XCas9 and Cas9-NG Hybrid","volume":"62","author":"Niu","year":"2020","journal-title":"J. Integr. Plant Biol."},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1111\/pbi.12025","article-title":"Fortifying Plants with the Essential Amino Acids Lysine and Methionine to Improve Nutritional Quality","volume":"11","author":"Galili","year":"2013","journal-title":"Plant Biotechnol. J."},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1007\/BF00022530","article-title":"Commercialization of a Tomato with an Antisense Polygalacturonase Gene: The FLAVR SAVRTM Tomato Story","volume":"79","author":"Kramer","year":"1994","journal-title":"Euphytica"},{"key":"ref_109","doi-asserted-by":"crossref","first-page":"koae316","DOI":"10.1093\/plcell\/koae316","article-title":"Efficient in Situ Epitope Tagging of Rice Genes by Nuclease-Mediated Prime Editing","volume":"37","author":"Li","year":"2025","journal-title":"Plant Cell"},{"key":"ref_110","doi-asserted-by":"crossref","unstructured":"Bhuyan, S.J., Kumar, M., Ramrao Devde, P., Rai, A.C., Mishra, A.K., Singh, P.K., and Siddique, K.H.M. (2023). Progress in Gene Editing Tools, Implications and Success in Plants: A Review. Front. Genome Ed., 5.","DOI":"10.3389\/fgeed.2023.1272678"},{"key":"ref_111","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1111\/pbi.14233","article-title":"Genome Editing of a Dominant Resistance Gene for Broad-spectrum Resistance to Bacterial Diseases in Rice without Growth Penalty","volume":"22","author":"Wang","year":"2024","journal-title":"Plant Biotechnol. J."},{"key":"ref_112","doi-asserted-by":"crossref","unstructured":"Illa-Berenguer, E., LaFayette, P.R., and Parrott, W.A. (2023). Editing Efficiencies with Cas9 Orthologs, Cas12a Endonucleases, and Temperature in Rice. Front. Genome Ed., 5.","DOI":"10.3389\/fgeed.2023.1074641"},{"key":"ref_113","doi-asserted-by":"crossref","first-page":"e70133","DOI":"10.1002\/pei3.70133","article-title":"CRISPR\/Cas Genome Editing and Its Applications in Cereal Crop Improvement","volume":"7","author":"Kaniganti","year":"2026","journal-title":"Plant-Environ. Interact."},{"key":"ref_114","doi-asserted-by":"crossref","first-page":"2008","DOI":"10.1093\/mp\/sst121","article-title":"Application of the CRISPR\u2013Cas System for Efficient Genome Engineering in Plants","volume":"6","author":"Mao","year":"2013","journal-title":"Mol. Plant"},{"key":"ref_115","doi-asserted-by":"crossref","first-page":"860","DOI":"10.1111\/pbi.14544","article-title":"Engineering a Robust Cas12i3 Variant-mediated Wheat Genome Editing System","volume":"23","author":"Wang","year":"2025","journal-title":"Plant Biotechnol. J."},{"key":"ref_116","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/21645698.2023.2256930","article-title":"CRISPR\/Cas9-Gene Editing Approaches in Plant Breeding","volume":"14","author":"Saini","year":"2023","journal-title":"GM Crops Food"},{"key":"ref_117","doi-asserted-by":"crossref","unstructured":"Jaganathan, D., Ramasamy, K., Sellamuthu, G., Jayabalan, S., and Venkataraman, G. (2018). CRISPR for Crop Improvement: An Update Review. Front. Plant Sci., 9.","DOI":"10.3389\/fpls.2018.00985"},{"key":"ref_118","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1038\/s41579-019-0299-x","article-title":"Evolutionary Classification of CRISPR\u2013Cas Systems: A Burst of Class 2 and Derived Variants","volume":"18","author":"Makarova","year":"2020","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_119","doi-asserted-by":"crossref","unstructured":"Ge, F., Peng, C., Du, Y., Chen, Y., Zhao, Z., Yu, M., Feng, H., Xie, Y., Sun, S., and Liu, S. (2026). Synergistic Engineering of Cas\u03b4 Nuclease for Robust Genome Editing. J. Integr. Plant Biol.","DOI":"10.1111\/jipb.70222"},{"key":"ref_120","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1038\/s41422-024-01050-y","article-title":"Structure and Genome Editing Activity of the Novel CRISPR-Cas12o1 Effector","volume":"35","author":"Duan","year":"2024","journal-title":"Cell Res."},{"key":"ref_121","doi-asserted-by":"crossref","first-page":"gkaf1358","DOI":"10.1093\/nar\/gkaf1358","article-title":"Cas\u03b4, an Evolutionary Transitional CRISPR System Enables Efficient Genome Editing across Animals and Plants","volume":"53","author":"Yang","year":"2025","journal-title":"Nucleic Acids Res."},{"key":"ref_122","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1111\/pbi.14574","article-title":"Engineering an Optimized Hypercompact CRISPR\/Cas12j-8 System for Efficient Genome Editing in Plants","volume":"23","author":"Bai","year":"2025","journal-title":"Plant Biotechnol. J."},{"key":"ref_123","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1126\/science.abb1400","article-title":"CRISPR-Cas\u03a6 from Huge Phages Is a Hypercompact Genome Editor","volume":"369","author":"Pausch","year":"2020","journal-title":"Science"},{"key":"ref_124","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1007\/s11248-026-00481-0","article-title":"Application of Compact CRISPR\/Cas Nucleases for Citrus Genome Editing","volume":"35","author":"Zhu","year":"2026","journal-title":"Transgenic Res."},{"key":"ref_125","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1038\/s41422-022-00771-2","article-title":"The Compact Cas\u03c0 (Cas12l) \u2018Bracelet\u2019 Provides a Unique Structural Platform for DNA Manipulation","volume":"33","author":"Sun","year":"2023","journal-title":"Cell Res."},{"key":"ref_126","doi-asserted-by":"crossref","first-page":"3478","DOI":"10.1038\/s41467-024-47800-9","article-title":"CoCas9 Is a Compact Nuclease from the Human Microbiome for Efficient and Precise Genome Editing","volume":"15","author":"Pedrazzoli","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_127","doi-asserted-by":"crossref","unstructured":"Sukegawa, S., Nureki, O., Toki, S., and Saika, H. (2023). Genome Editing in Rice Mediated by Miniature Size Cas Nuclease SpCas12f. Front. Genome Ed., 5.","DOI":"10.3389\/fgeed.2023.1138843"},{"key":"ref_128","doi-asserted-by":"crossref","first-page":"6102","DOI":"10.1038\/s41467-023-41802-9","article-title":"Efficient Plant Genome Engineering Using a Probiotic Sourced CRISPR-Cas9 System","volume":"14","author":"Zhong","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_129","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1007\/s42994-021-00051-4","article-title":"Expanding the Range of CRISPR\/Cas9-Directed Genome Editing in Soybean","volume":"3","author":"He","year":"2022","journal-title":"aBIOTECH"},{"key":"ref_130","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1186\/s13059-023-02929-6","article-title":"Boosting Genome Editing Efficiency in Human Cells and Plants with Novel LbCas12a Variants","volume":"24","author":"Zhang","year":"2023","journal-title":"Genome Biol."},{"key":"ref_131","doi-asserted-by":"crossref","first-page":"3164","DOI":"10.1111\/pbi.14438","article-title":"PhieDBEs: A DBD-containing, PAM-flexible, High-efficiency Dual Base Editor Toolbox with Wide Targeting Scope for Use in Plants","volume":"22","author":"Zheng","year":"2024","journal-title":"Plant Biotechnol. J."},{"key":"ref_132","doi-asserted-by":"crossref","first-page":"934","DOI":"10.1111\/pbi.13774","article-title":"PhieABEs: A PAM-less\/Free High-efficiency Adenine Base Editor Toolbox with Wide Target Scope in Plants","volume":"20","author":"Tan","year":"2022","journal-title":"Plant Biotechnol. J."},{"key":"ref_133","doi-asserted-by":"crossref","unstructured":"Sretenovic, S., Liu, S., Li, G., Cheng, Y., Fan, T., Xu, Y., Zhou, J., Zheng, X., Coleman, G., and Zhang, Y. (2021). Exploring C-To-G Base Editing in Rice, Tomato, and Poplar. Front. Genome Ed., 3.","DOI":"10.3389\/fgeed.2021.756766"},{"key":"ref_134","doi-asserted-by":"crossref","unstructured":"Gallo, D., Meunier, A.-C., and P\u00e9rin, C. (2025). A Long Journey towards Genome Editing Technologies in Plants: A Technical and Critical Review of Genome Editing Technologies. Front. Genome Ed., 7.","DOI":"10.3389\/fgeed.2025.1663352"},{"key":"ref_135","doi-asserted-by":"crossref","unstructured":"Westberg, I., Carlsen, F.M., Johansen, I.E., and Petersen, B.L. (2023). Cytosine Base Editors Optimized for Genome Editing in Potato Protoplasts. Front. Genome Ed., 5.","DOI":"10.3389\/fgeed.2023.1247702"},{"key":"ref_136","doi-asserted-by":"crossref","unstructured":"Achary, V.M.M., Syombua, E.D., Kaur, S., Katamreddy, S.C., Zou, D., Hearne, S.J., and Bandyopadhyay, A. (2026). Advances in the Use of Morphogenic Regulators and Peptide Regenerating Factors for Boosting Plant Transformation and Genome Editing. Front. Plant Sci., 17.","DOI":"10.3389\/fpls.2026.1699984"},{"key":"ref_137","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1007\/s00299-026-03820-x","article-title":"Evaluation of Computational Tools for the Prediction of CRISPR\/SpCas9 GRNA Activity in Plants","volume":"45","author":"Gong","year":"2026","journal-title":"Plant Cell Rep."},{"key":"ref_138","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1007\/s42994-024-00157-5","article-title":"Developing a CRISPR\/FrCas9 System for Core Promoter Editing in Rice","volume":"5","author":"Wang","year":"2024","journal-title":"aBIOTECH"},{"key":"ref_139","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1007\/s42994-024-00177-1","article-title":"Prime Editing Enables Precise Genome Modification of a Populus Hybrid","volume":"5","author":"Zou","year":"2024","journal-title":"aBIOTECH"},{"key":"ref_140","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1186\/s13007-023-01010-4","article-title":"Targeted Gene Deletion with SpCas9 and Multiple Guide RNAs in Arabidopsis Thaliana: Four Are Better than Two","volume":"19","author":"Ordon","year":"2023","journal-title":"Plant Methods"},{"key":"ref_141","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Cai, Y., Sun, S., Han, T., Chen, L., and Hou, W. (2022). Using Staphylococcus Aureus Cas9 to Expand the Scope of Potential Gene Targets for Genome Editing in Soybean. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232112789"},{"key":"ref_142","doi-asserted-by":"crossref","unstructured":"Sony, S.K., Kaul, T., Motelb, K.F.A., Thangaraj, A., Bharti, J., Kaul, R., Verma, R., and Nehra, M. (2023). CRISPR\/Cas9-mediated Homology Donor Repair Base Editing Confers Glyphosate Resistance to Rice (Oryza sativa L.). Front. Plant Sci., 14.","DOI":"10.3389\/fpls.2023.1122926"},{"key":"ref_143","doi-asserted-by":"crossref","unstructured":"Cui, Y., Zhao, J., Gao, Y., Zhao, R., Zhang, J., and Kong, L. (2021). Efficient Multi-Sites Genome Editing and Plant Regeneration via Somatic Embryogenesis in Picea Glauca. Front. Plant Sci., 12.","DOI":"10.3389\/fpls.2021.751891"},{"key":"ref_144","doi-asserted-by":"crossref","first-page":"100778","DOI":"10.1016\/j.xplc.2023.100778","article-title":"Population Genomic Analysis Unravels the Evolutionary Roadmap of Pericarp Color in Rice","volume":"5","author":"Xie","year":"2024","journal-title":"Plant Commun."},{"key":"ref_145","doi-asserted-by":"crossref","first-page":"100026","DOI":"10.1016\/j.abiote.2026.100026","article-title":"Enhancing the Efficiency of Nuclease-Based Prime Editing in Rice with the Tf1 Reverse Transcriptase","volume":"7","author":"Ma","year":"2026","journal-title":"aBIOTECH"},{"key":"ref_146","doi-asserted-by":"crossref","first-page":"911","DOI":"10.14348\/molcells.2021.0237","article-title":"RPS5A Promoter-Driven Cas9 Produces Heritable Virus-Induced Genome Editing in Nicotiana attenuata","volume":"44","author":"Oh","year":"2021","journal-title":"Mol. Cells"},{"key":"ref_147","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1094\/MPMI-10-24-0127-TA","article-title":"Generation of Inheritable A-to-G Transitions Using Adenine Base Editing and NG-PAM Cas9 in Arabidopsis thaliana","volume":"38","author":"Tan","year":"2025","journal-title":"Mol. Plant-Microbe Interact."},{"key":"ref_148","doi-asserted-by":"crossref","first-page":"1743","DOI":"10.1093\/pcp\/pcae100","article-title":"Heritable Tissue-Culture-Free Gene Editing in Nicotiana Benthamiana through Viral Delivery of SpCas9 and SgRNA","volume":"65","author":"Yoshida","year":"2024","journal-title":"Plant Cell Physiol."},{"key":"ref_149","doi-asserted-by":"crossref","unstructured":"Qaim, M., and Kouser, S. (2013). Genetically Modified Crops and Food Security. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0064879"},{"key":"ref_150","doi-asserted-by":"crossref","unstructured":"Kl\u00fcmper, W., and Qaim, M. (2014). A Meta-Analysis of the Impacts of Genetically Modified Crops. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0111629"},{"key":"ref_151","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/j.tplants.2015.02.004","article-title":"Plant Synthetic Biology","volume":"20","author":"Liu","year":"2015","journal-title":"Trends Plant Sci."},{"key":"ref_152","doi-asserted-by":"crossref","first-page":"1428","DOI":"10.21474\/IJAR01\/3650","article-title":"Linseed: A Valuable Crop Plant","volume":"5","author":"Dash","year":"2017","journal-title":"Int. J. Adv. Res."},{"key":"ref_153","unstructured":"WorldFloraDB (2026, April 20). Available online: https:\/\/www.worldfloradb.net\/plants\/linum-usitatissimum-var-usitatissimum."},{"key":"ref_154","doi-asserted-by":"crossref","unstructured":"Soto-Cerda, B.J., Diederichsen, A., Ragupathy, R., and Cloutier, S. (2013). Genetic Characterization of a Core Collection of Flax (Linum Usitatissimum L.) Suitable for Association Mapping Studies and Evidence of Divergent Selection between Fiber and Linseed Types. BMC Plant Biol., 13.","DOI":"10.1186\/1471-2229-13-78"},{"key":"ref_155","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.crvi.2016.12.002","article-title":"SSR and Morphological Trait Based Population Structure Analysis of 130 Diverse Flax (Linum usitatissimum L.) Accessions","volume":"340","author":"Choudhary","year":"2017","journal-title":"C. R. Biol."},{"key":"ref_156","doi-asserted-by":"crossref","unstructured":"Xie, D., Dai, Z., Yang, Z., Tang, Q., Sun, J., Yang, X., Song, X., Lu, Y., Zhao, D., and Zhang, L. (2018). Genomic Variations and Association Study of Agronomic Traits in Flax. BMC Genom., 19.","DOI":"10.1186\/s12864-018-4899-z"},{"key":"ref_157","doi-asserted-by":"crossref","unstructured":"You, F.M., Xiao, J., Li, P., Yao, Z., Jia, G., He, L., Kumar, S., Soto-Cerda, B., Duguid, S.D., and Booker, H.M. (2018). Genome-Wide Association Study and Selection Signatures Detect Genomic Regions Associated with Seed Yield and Oil Quality in Flax. Int. J. Mol. Sci., 19.","DOI":"10.20944\/preprints201807.0397.v2"},{"key":"ref_158","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1007\/s00122-012-1860-4","article-title":"Simple Sequence Repeat Marker Development from Bacterial Artificial Chromosome End Sequences and Expressed Sequence Tags of Flax (Linum usitatissimum L.)","volume":"125","author":"Cloutier","year":"2012","journal-title":"Theor. Appl. Genet."},{"key":"ref_159","doi-asserted-by":"crossref","first-page":"1783","DOI":"10.1007\/s00122-012-1953-0","article-title":"Integrated Consensus Genetic and Physical Maps of Flax (Linum usitatissimum L.)","volume":"125","author":"Cloutier","year":"2012","journal-title":"Theor. Appl. Genet."},{"key":"ref_160","doi-asserted-by":"crossref","unstructured":"Kumar, S., You, F.M., and Cloutier, S. (2012). Genome Wide SNP Discovery in Flax through next Generation Sequencing of Reduced Representation Libraries. BMC Genom., 13.","DOI":"10.1186\/1471-2164-13-684"},{"key":"ref_161","doi-asserted-by":"crossref","unstructured":"Povkhova, L.V., Melnikova, N.V., Rozhmina, T.A., Novakovskiy, R.O., Pushkova, E.N., Dvorianinova, E.M., Zhuchenko, A.A., Kamionskaya, A.M., Krasnov, G.S., and Dmitriev, A.A. (2021). Genes Associated with the Flax Plant Type (Oil or Fiber) Identified Based on Genome and Transcriptome Sequencing Data. Plants, 10.","DOI":"10.3390\/plants10122616"},{"key":"ref_162","doi-asserted-by":"crossref","unstructured":"Sa, R., Yi, L., Siqin, B., An, M., Bao, H., Song, X., Wang, S., Li, Z., Zhang, Z., and Hazaisi, H. (2021). Chromosome-Level Genome Assembly and Annotation of the Fiber Flax (Linum usitatissimum) Genome. Front. Genet., 12.","DOI":"10.3389\/fgene.2021.735690"},{"key":"ref_163","doi-asserted-by":"crossref","unstructured":"Zhao, X., Yi, L., Zuo, Y., Gao, F., Cheng, Y., Zhang, H., Zhou, Y., Jia, X., Su, S., and Zhang, D. (2023). High-Quality Genome Assembly and Genome-Wide Association Study of Male Sterility Provide Resources for Flax Improvement. Plants, 12.","DOI":"10.3390\/plants12152773"},{"key":"ref_164","doi-asserted-by":"crossref","unstructured":"Wang, N., Qi, F., Wang, F., Lin, Y., Xiaoyang, C., Peng, Z., Zhang, B., Qi, X., Deyholos, M.K., and Zhang, J. (2023). Evaluation of Differentially Expressed Genes in Leaves vs. Roots Subjected to Drought Stress in Flax (Linum usitatissimum L.). Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241512019"},{"key":"ref_165","doi-asserted-by":"crossref","first-page":"105019","DOI":"10.1016\/j.envexpbot.2022.105019","article-title":"Integrating Multi-Locus Genome-Wide Association Studies with Transcriptomic Data to Identify Genetic Loci Underlying Adult Root Trait Responses to Drought Stress in Flax (Linum usitatissimum L.)","volume":"202","author":"Larama","year":"2022","journal-title":"Environ. Exp. Bot."},{"key":"ref_166","doi-asserted-by":"crossref","unstructured":"Wang, W., Wang, L., Wang, L., Tan, M., Ogutu, C.O., Yin, Z., Zhou, J., Wang, J., Wang, L., and Yan, X. (2021). Transcriptome Analysis and Molecular Mechanism of Linseed (Linum usitatissimum L.) Drought Tolerance under Repeated Drought Using Single-Molecule Long-Read Sequencing. BMC Genom., 22.","DOI":"10.1186\/s12864-021-07416-5"},{"key":"ref_167","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1104\/pp.105.064451","article-title":"Two FAD3 Desaturase Genes Control the Level of Linolenic Acid in Flax Seed","volume":"139","author":"Vrinten","year":"2005","journal-title":"Plant Physiol."},{"key":"ref_168","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1007\/BF00269571","article-title":"Genetic Transformation of Flax (Linum usitatissimum) by Agrobacterium Tumefaciens: Regeneration of Transformed Shoots via a Callus Phase","volume":"6","author":"Basiran","year":"1987","journal-title":"Plant Cell Rep."},{"key":"ref_169","doi-asserted-by":"crossref","unstructured":"Bastaki, N.K., and Cullis, C.A. (2014). Floral-Dip Transformation of Flax (Linum usitatissimum) to Generate Transgenic Progenies with a High Transformation Rate. J. Vis. Exp., e52189.","DOI":"10.3791\/52189"},{"key":"ref_170","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1007\/s11240-007-9335-z","article-title":"Sonication Assisted Agrobacterium-Mediated Transformation Enhances the Transformation Efficiency in Flax (Linum usitatissimum L.)","volume":"94","year":"2008","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_171","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/0168-9452(93)90137-O","article-title":"Transgenic Flax Plants from Agrobacterium Mediated Transformation: Incidence of Chimeric Regenerants and Inheritance of Transgenic Plants","volume":"91","author":"Dong","year":"1993","journal-title":"Plant Sci."},{"key":"ref_172","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1007\/BF00017455","article-title":"Regeneration of Flax Plants Transformed by Agrobacterium Rhizogenes","volume":"11","author":"Zhan","year":"1988","journal-title":"Plant Mol. Biol."},{"key":"ref_173","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1186\/s13007-025-01401-9","article-title":"CRISPR-Cas13a as a next-Generation Tool for Rapid and Precise Plant RNA Virus Diagnostics","volume":"21","author":"Karimi","year":"2025","journal-title":"Plant Methods"},{"key":"ref_174","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/S0176-1617(00)80055-3","article-title":"Shoots and Embryo-like Structures Regenerated from Cultured Flax (Linum usitatissimum L.) Hypocotyl Segments","volume":"157","author":"Obert","year":"2000","journal-title":"J. Plant Physiol."},{"key":"ref_175","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1007\/s00299-003-0662-1","article-title":"Regeneration of Flax (Linum usitatissimum L.) Plants from Anther Culture and Somatic Tissue with Increased Resistance to Fusarium Oxysporum","volume":"22","author":"Mankowska","year":"2003","journal-title":"Plant Cell Rep."},{"key":"ref_176","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1016\/j.jplph.2004.06.002","article-title":"Moderation of Morphogenetic and Oxidative Stress Responses in Flax in Vitro Cultures by Hydroxynonenal and Desferrioxamine","volume":"162","author":"Obert","year":"2005","journal-title":"J. Plant Physiol."},{"key":"ref_177","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1007\/s12892-024-00265-0","article-title":"Stimulating Endogenous Hormone Content by Plant Extracts: Increased in Vitro Regeneration of Flax (Linum Usitatissimum) Cultivars","volume":"28","author":"Oguz","year":"2025","journal-title":"J. Crop Sci. Biotechnol."},{"key":"ref_178","first-page":"358","article-title":"Efficient Plantlet Regeneration from Hypocotyl Explants and Agrobacterium-Mediated Genetic Transformation of Flax (Linum usitatissimum L.)","volume":"19","author":"Mandal","year":"2018","journal-title":"Plant Cell Biotechnol. Mol. Biol."},{"key":"ref_179","first-page":"120","article-title":"Improved Flax Regeneration from Hypocotyls Using Thidiazuron as a Cytokinin Source","volume":"14","author":"Bretagne","year":"1994","journal-title":"Plant Cell Rep."},{"key":"ref_180","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1016\/j.biotechadv.2009.02.004","article-title":"Doubled Haploid Production in Flax (Linum usitatissimum L.)","volume":"27","author":"Obert","year":"2009","journal-title":"Biotechnol. Adv."},{"key":"ref_181","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1016\/S0176-1617(97)80015-6","article-title":"Transformation in Protoplast Cultures of Linum Usitatissimum and L. Suffruticosum Mediated with PEG and with Agrobacterium Tumefaciens","volume":"151","author":"Ling","year":"1997","journal-title":"J. Plant Physiol."},{"key":"ref_182","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1007\/BF00233320","article-title":"High Efficiency Agrobacterium-Mediated Gene Transfer to Flax","volume":"13","author":"Bauer","year":"1994","journal-title":"Plant Cell Rep."},{"key":"ref_183","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1007\/s00299-006-0280-9","article-title":"The Use of the Phosphomannose Isomerase Gene as Alternative Selectable Marker for Agrobacterium-Mediated Transformation of Flax (Linum usitatissimum)","volume":"26","author":"Lamblin","year":"2007","journal-title":"Plant Cell Rep."},{"key":"ref_184","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1007\/s00709-010-0261-2","article-title":"ER Disruption and GFP Degradation during Non-Regenerable Transformation of Flax with Agrobacterium Tumefaciens","volume":"249","author":"Bleho","year":"2012","journal-title":"Protoplasma"},{"key":"ref_185","doi-asserted-by":"crossref","unstructured":"Li, F., Xue, M., Guo, D., Zhu, L., Li, Y., and Xie, L. (2025). A Truncated Endogenous U6 Promoter Enables High-Efficiency CRISPR Editing in Flax (Linum usitatissimum L.). Plants, 14.","DOI":"10.3390\/plants14071142"},{"key":"ref_186","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1263\/jbb.105.636","article-title":"Enrichment of Carotenoids in Flaxseed (Linum usitatissimum) by Metabolic Engineering with Introduction of Bacterial Phytoene Synthase Gene CrtB","volume":"105","author":"Fujisawa","year":"2008","journal-title":"J. Biosci. Bioeng."},{"key":"ref_187","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1007\/s11103-018-0725-x","article-title":"A Genome-Wide Analysis of the Flax (Linum usitatissimum L.) Dirigent Protein Family: From Gene Identification and Evolution to Differential Regulation","volume":"97","author":"Corbin","year":"2018","journal-title":"Plant Mol. Biol."},{"key":"ref_188","doi-asserted-by":"crossref","unstructured":"Wang, Y., Qi, Y., Wang, L., Xu, C., Li, W., Dang, Z., Zhao, W., Wang, P., Xie, Y., and Niu, Y. (2025). Genome-Wide Identification and Expression Profiling of Dehydration-Responsive Element-Binding Family Genes in Flax (Linum usitatissimum L.). Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26073074"},{"key":"ref_189","doi-asserted-by":"crossref","unstructured":"Cullis, C.A. (2019). Genetics and Genomics of Linum, Springer International Publishing.","DOI":"10.1007\/978-3-030-23964-0"},{"key":"ref_190","doi-asserted-by":"crossref","first-page":"810","DOI":"10.1038\/313810a0","article-title":"Identification of DNA Sequences Required for Activity of the Cauliflower Mosaic Virus 35S Promoter","volume":"313","author":"Odell","year":"1985","journal-title":"Nature"},{"key":"ref_191","doi-asserted-by":"crossref","unstructured":"Chantreau, M., Grec, S., Gutierrez, L., Dalmais, M., Pineau, C., Demailly, H., Paysant-Leroux, C., Tavernier, R., Trouv\u00e9, J.-P., and Chatterjee, M. (2013). PT-Flax (Phenotyping and TILLinG of Flax): Development of a Flax (Linum Usitatissimum L.) Mutant Population and TILLinG Platform for Forward and Reverse Genetics. BMC Plant Biol., 13.","DOI":"10.1186\/1471-2229-13-159"},{"key":"ref_192","doi-asserted-by":"crossref","unstructured":"Preisner, M., Kulma, A., Zebrowski, J., Dymi\u0144ska, L., Hanuza, J., Arendt, M., Starzycki, M., and Szopa, J. (2014). Manipulating Cinnamyl Alcohol Dehydrogenase (CAD) Expression in Flax Affects Fibre Composition and Properties. BMC Plant Biol., 14.","DOI":"10.1186\/1471-2229-14-50"},{"key":"ref_193","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.plaphy.2008.09.011","article-title":"Caffeoyl Coenzyme A O-Methyltransferase down-Regulation Is Associated with Modifications in Lignin and Cell-Wall Architecture in Flax Secondary Xylem","volume":"47","author":"Day","year":"2009","journal-title":"Plant Physiol. Biochem."},{"key":"ref_194","doi-asserted-by":"crossref","first-page":"794","DOI":"10.1111\/j.1365-313X.2008.03444.x","article-title":"The Roles of the Cytoskeleton during Cellulose Deposition at the Secondary Cell Wall","volume":"54","author":"Wightman","year":"2008","journal-title":"Plant J."},{"key":"ref_195","doi-asserted-by":"crossref","first-page":"1065","DOI":"10.1002\/cbin.10837","article-title":"Expression Analysis of Cellulose Synthase and Main Cytoskeletal Protein Genes in Flax (Linum usitatissimum L.)","volume":"43","author":"Galinousky","year":"2019","journal-title":"Cell Biol. Int."},{"key":"ref_196","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1111\/pbi.12350","article-title":"Functional Analyses of Cellulose Synthase Genes in Flax (Linum usitatissimum) by Virus-induced Gene Silencing","volume":"13","author":"Chantreau","year":"2015","journal-title":"Plant Biotechnol. J."},{"key":"ref_197","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1038\/nature17946","article-title":"Programmable Editing of a Target Base in Genomic DNA without Double-Stranded DNA Cleavage","volume":"533","author":"Komor","year":"2016","journal-title":"Nature"},{"key":"ref_198","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1038\/s41586-019-1711-4","article-title":"Search-and-Replace Genome Editing without Double-Strand Breaks or Donor DNA","volume":"576","author":"Anzalone","year":"2019","journal-title":"Nature"},{"key":"ref_199","doi-asserted-by":"crossref","unstructured":"Li, Y., Wu, X., Zhang, Y., and Zhang, Q. (2022). CRISPR\/Cas Genome Editing Improves Abiotic and Biotic Stress Tolerance of Crops. Front. Genome Ed., 4.","DOI":"10.3389\/fgeed.2022.987817"},{"key":"ref_200","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1186\/s13059-020-02204-y","article-title":"Engineering Crops of the Future: CRISPR Approaches to Develop Climate-Resilient and Disease-Resistant Plants","volume":"21","author":"Zaidi","year":"2020","journal-title":"Genome Biol."},{"key":"ref_201","doi-asserted-by":"crossref","unstructured":"Metje-Sprink, J., Menz, J., Modrzejewski, D., and Sprink, T. (2019). DNA-Free Genome Editing: Past, Present and Future. Front. Plant Sci., 9.","DOI":"10.3389\/fpls.2018.01957"},{"key":"ref_202","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1007\/s10142-019-00667-2","article-title":"Flax Tubulin and CesA Superfamilies Represent Attractive and Challenging Targets for a Variety of Genome- and Base-Editing Applications","volume":"20","author":"Morello","year":"2020","journal-title":"Funct. Integr. Genom."},{"key":"ref_203","doi-asserted-by":"crossref","first-page":"1651","DOI":"10.1111\/pbi.13383","article-title":"Genome Editing with the CRISPR-Cas System: An Art, Ethics and Global Regulatory Perspective","volume":"18","author":"Zhang","year":"2020","journal-title":"Plant Biotechnol. J."},{"key":"ref_204","doi-asserted-by":"crossref","unstructured":"Liu, C., Zhang, T., Yang, X., Wang, L., Long, Y., Hasi, A., and Pei, X. (2023). A LuALS Mutation with High Sulfonylurea Herbicide Resistance in Linum usitatissimum L. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24032820"},{"key":"ref_205","first-page":"60","article-title":"The Impact of Genetic Engineering on Maize Herbicide Tolerance","volume":"15","author":"Wu","year":"2024","journal-title":"Maize Genom. Genet."},{"key":"ref_206","doi-asserted-by":"crossref","unstructured":"Siddique, S. (2022). Role of CRISPR\/Cas9 in Soybean (Glycine max L.) Quality Improvement. Soybean\u2014Recent Advances in Research and Applications, IntechOpen.","DOI":"10.5772\/intechopen.102812"},{"key":"ref_207","doi-asserted-by":"crossref","unstructured":"Dvorianinova, E.M., Zinovieva, O.L., Pushkova, E.N., Zhernova, D.A., Rozhmina, T.A., Povkhova, L.V., Novakovskiy, R.O., Sigova, E.A., Turba, A.A., and Borkhert, E.V. (2023). Key FAD2, FAD3, and SAD Genes Involved in the Fatty Acid Synthesis in Flax Identified Based on Genomic and Transcriptomic Data. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241914885"},{"key":"ref_208","doi-asserted-by":"crossref","first-page":"3518","DOI":"10.1093\/jxb\/erad096","article-title":"CRISPR\/Cas9 and Agrobacterium tumefaciens Virulence Proteins Synergistically Increase Efficiency of Precise Genome Editing via Homology Directed Repair in Plants","volume":"74","author":"Tang","year":"2023","journal-title":"J. Exp. Bot."},{"key":"ref_209","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1007\/978-1-0716-1323-8_10","article-title":"RNAi-Based Gene Silencing in Sugarcane for Production of Biofuel","volume":"2290","author":"Murugan","year":"2021","journal-title":"Methods Mol. Biol."},{"key":"ref_210","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1016\/j.cj.2021.08.006","article-title":"Generation of a High-Efficiency Adenine Base Editor with TadA8e for Developing Wheat Dinitroaniline-Resistant Germplasm","volume":"10","author":"Han","year":"2022","journal-title":"Crop J."},{"key":"ref_211","doi-asserted-by":"crossref","first-page":"1332","DOI":"10.1007\/s11434-015-0848-2","article-title":"A Detailed Procedure for CRISPR\/Cas9-Mediated Gene Editing in Arabidopsis Thaliana","volume":"60","author":"Liu","year":"2015","journal-title":"Sci. Bull."},{"key":"ref_212","doi-asserted-by":"crossref","first-page":"1780","DOI":"10.1093\/plphys\/kiac037","article-title":"An Update on Precision Genome Editing by Homology-Directed Repair in Plants","volume":"188","author":"Chen","year":"2022","journal-title":"Plant Physiol."},{"key":"ref_213","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1186\/s13578-017-0148-4","article-title":"Plant Genome Editing with TALEN and CRISPR","volume":"7","author":"Malzahn","year":"2017","journal-title":"Cell Biosci."},{"key":"ref_214","doi-asserted-by":"crossref","first-page":"1139","DOI":"10.1111\/tpj.12704","article-title":"The CRISPR\/Cas System Can Be Used as Nuclease for in Planta Gene Targeting and as Paired Nickases for Directed Mutagenesis in Arabidopsis Resulting in Heritable Progeny","volume":"80","author":"Schiml","year":"2014","journal-title":"Plant J."},{"key":"ref_215","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1007\/s11103-015-0342-x","article-title":"Comparison of CRISPR\/Cas9 Expression Constructs for Efficient Targeted Mutagenesis in Rice","volume":"88","author":"Mikami","year":"2015","journal-title":"Plant Mol. Biol."},{"key":"ref_216","doi-asserted-by":"crossref","first-page":"1917","DOI":"10.1104\/pp.15.01696","article-title":"Oligonucleotide-Mediated Genome Editing Provides Precision and Function to Engineered Nucleases and Antibiotics in Plants","volume":"170","author":"Sauer","year":"2016","journal-title":"Plant Physiol."},{"key":"ref_217","doi-asserted-by":"crossref","first-page":"1402","DOI":"10.1038\/s41587-020-0581-5","article-title":"Targeted, Efficient Sequence Insertion and Replacement in Rice","volume":"38","author":"Lu","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"ref_218","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1038\/s41477-021-00991-1","article-title":"Precise Plant Genome Editing Using Base Editors and Prime Editors","volume":"7","author":"Molla","year":"2021","journal-title":"Nat. Plants"},{"key":"ref_219","doi-asserted-by":"crossref","first-page":"1795","DOI":"10.1093\/plphys\/kiab591","article-title":"Improvement of Base Editors and Prime Editors Advances Precision Genome Engineering in Plants","volume":"188","author":"Hua","year":"2022","journal-title":"Plant Physiol."},{"key":"ref_220","doi-asserted-by":"crossref","first-page":"uhac047","DOI":"10.1093\/hr\/uhac047","article-title":"Genotype-Independent Plant Transformation","volume":"9","author":"Maren","year":"2022","journal-title":"Hortic. Res."},{"key":"ref_221","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1186\/s42397-023-00140-3","article-title":"Genome Editing in Cotton: Challenges and Opportunities","volume":"6","author":"Khan","year":"2023","journal-title":"J. Cotton Res."},{"key":"ref_222","doi-asserted-by":"crossref","first-page":"648","DOI":"10.1111\/pbi.12663","article-title":"Significant Enhancement of Fatty Acid Composition in Seeds of the Allohexaploid, Camelina Sativa, Using CRISPR\/Cas9 Gene Editing","volume":"15","author":"Jiang","year":"2017","journal-title":"Plant Biotechnol. J."},{"key":"ref_223","doi-asserted-by":"crossref","unstructured":"Ingvardsen, C.R., and Brinch-Pedersen, H. (2023). Challenges and Potentials of New Breeding Techniques in Cannabis Sativa. Front. Plant Sci., 14.","DOI":"10.3389\/fpls.2023.1154332"},{"key":"ref_224","doi-asserted-by":"crossref","unstructured":"Zhang, Y., and Showalter, A.M. (2020). CRISPR\/Cas9 Genome Editing Technology: A Valuable Tool for Understanding Plant Cell Wall Biosynthesis and Function. Front. Plant Sci., 11.","DOI":"10.3389\/fpls.2020.589517"},{"key":"ref_225","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1016\/j.tibtech.2013.04.004","article-title":"ZFN, TALEN, and CRISPR\/Cas-Based Methods for Genome Engineering","volume":"31","author":"Gaj","year":"2013","journal-title":"Trends Biotechnol."},{"key":"ref_226","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.jgg.2013.03.009","article-title":"TALENs: Customizable Molecular DNA Scissors for Genome Engineering of Plants","volume":"40","author":"Chen","year":"2013","journal-title":"J. Genet. Genom."},{"key":"ref_227","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1038\/nature02874","article-title":"RNA Silencing in Plants","volume":"431","author":"Baulcombe","year":"2004","journal-title":"Nature"},{"key":"ref_228","doi-asserted-by":"crossref","unstructured":"MacMillan, C.P., Birke, H., Chuah, A., Brill, E., Tsuji, Y., Ralph, J., Dennis, E.S., Llewellyn, D., and Pettolino, F.A. (2017). Tissue and Cell-Specific Transcriptomes in Cotton Reveal the Subtleties of Gene Regulation Underlying the Diversity of Plant Secondary Cell Walls. BMC Genom., 18, Erratum in BMC Genom. 2018, 19, 261. https:\/\/doi.org\/10.1186\/s12864-018-4634-9.","DOI":"10.1186\/s12864-017-3902-4"},{"key":"ref_229","doi-asserted-by":"crossref","first-page":"lsab033","DOI":"10.1093\/jlb\/lsab033","article-title":"Are Genome-Edited Micro-Organisms Covered by Directive 2009\/41\/EC?\u2014Implications of the CJEU\u2019s Judgment in the Case C-528\/16 for the Contained Use of Genome-Edited Micro-Organisms","volume":"9","author":"Dederer","year":"2022","journal-title":"J. Law Biosci."},{"key":"ref_230","first-page":"21","article-title":"Uropean Commission\u2019s Plans for a Special Regulation of Plants Created by New Genomic Techniques","volume":"2024","author":"Kahrmann","year":"2024","journal-title":"Eur. Pap."},{"key":"ref_231","doi-asserted-by":"crossref","unstructured":"Turnbull, C., Lillemo, M., and Hvoslef-Eide, T.A.K. (2021). Global Regulation of Genetically Modified Crops Amid the Gene Edited Crop Boom\u2014A Review. Front. Plant Sci., 12.","DOI":"10.3389\/fpls.2021.630396"},{"key":"ref_232","doi-asserted-by":"crossref","unstructured":"Normile, D. (ScienceInsider, 2019). Gene-edited foods are safe, Japanese panel concludes, ScienceInsider.","DOI":"10.1126\/science.aax3903"},{"key":"ref_233","doi-asserted-by":"crossref","first-page":"1281","DOI":"10.1038\/s41467-020-15053-x","article-title":"Multiplexed CRISPR Technologies for Gene Editing and Transcriptional Regulation","volume":"11","author":"McCarty","year":"2020","journal-title":"Nat. Commun."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/13\/6012\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,12]],"date-time":"2026-07-12T04:10:57Z","timestamp":1783829457000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/13\/6012"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,4]]},"references-count":233,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["ijms27136012"],"URL":"https:\/\/doi.org\/10.3390\/ijms27136012","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,4]]}}}