{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T19:51:40Z","timestamp":1790106700431,"version":"4.0.1"},"reference-count":40,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,5,17]],"date-time":"2025-05-17T00:00:00Z","timestamp":1747440000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Brazilian research agencies: National Council for Scientific and Technological Development (CNPq)","award":["470888\/2014-8"],"award-info":[{"award-number":["470888\/2014-8"]}]},{"name":"Brazilian research agencies: National Council for Scientific and Technological Development (CNPq)","award":["403587\/2021-2"],"award-info":[{"award-number":["403587\/2021-2"]}]},{"name":"Brazilian research agencies: National Council for Scientific and Technological Development (CNPq)","award":["141742\/2014-3"],"award-info":[{"award-number":["141742\/2014-3"]}]},{"name":"Brazilian research agencies: National Council for Scientific and Technological Development (CNPq)","award":["20160482"],"award-info":[{"award-number":["20160482"]}]},{"name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior-Brazil (CAPES)-Finance Code 001 and FIPE\/HCPA","award":["470888\/2014-8"],"award-info":[{"award-number":["470888\/2014-8"]}]},{"name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior-Brazil (CAPES)-Finance Code 001 and FIPE\/HCPA","award":["403587\/2021-2"],"award-info":[{"award-number":["403587\/2021-2"]}]},{"name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior-Brazil (CAPES)-Finance Code 001 and FIPE\/HCPA","award":["141742\/2014-3"],"award-info":[{"award-number":["141742\/2014-3"]}]},{"name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior-Brazil (CAPES)-Finance Code 001 and FIPE\/HCPA","award":["20160482"],"award-info":[{"award-number":["20160482"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Background: Genome editing at specific loci is an innovative therapeutic approach; however, it faces many challenges, so optimizing delivery vectors is essential to enhance the safety and efficacy of the CRISPR\/Cas9 system. This study investigated whether the hydrodynamic administration of liposomal CRISPR\/Cas9 complexes (LCs) in newborn mice induces off-target events or tumors. Methods: Liposomes were obtained through microfluidization. The CRISPR\/Cas9 plasmid and a donor plasmid containing the Idua cDNA (alpha-L-iduronidase enzyme) were incorporated by adsorption, and complexes (LCs) were characterized regarding physicochemical properties. C57BL\/6 newborn mice were divided in two groups, one received the complexes through hydrodynamic intravenous injection (n = 15) and the other was used as control (n = 15). After 21 months, mice were euthanized and organs were analyzed regarding histological characteristics. Lungs and liver were analyzed by qPCR searching for potential off-target sites in chromosomes 2, 5, 11, and 17 and on-target site in chromosome 6, identified by COSMID. Sequences were analyzed using an ICE tool for indels detection. Results: LCs exhibited 136 nm mean vesicle diameter with PDI below 0.15 and a zeta potential around +43 mV. Immediate biodistribution was predominant in the lungs and liver. There was no significant increase in tumor induction (20% in LCs vs. 33% in control). Molecular analyses indicated 0% off-target effects and around 3% on-target events. Conclusions: We conclude that this set of experiments demonstrates the potential of the chosen gRNA sequence to perform safe gene editing at the murine ROSA26 locus, corroborating the safety of the CRISPR\/Cas9 gene editing platform.<\/jats:p>","DOI":"10.3390\/pharmaceutics17050656","type":"journal-article","created":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T04:48:18Z","timestamp":1747716498000},"page":"656","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Newborn Intravenous Injection of Liposomal CRISPR\/Cas9 Complex Has No Incidence of Off-Targets or Tumors in Mice"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-7361-1770","authenticated-orcid":false,"given":"Vin\u00edcius","family":"Monteagudo","sequence":"first","affiliation":[{"name":"Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, Avenida Ipiranga, 2752\/lab 606, Porto Alegre 90610-000, RS, Brazil"},{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Larissa Cristina Barbosa","family":"Flores","sequence":"additional","affiliation":[{"name":"Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, Avenida Ipiranga, 2752\/lab 606, Porto Alegre 90610-000, RS, Brazil"},{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melaine","family":"Lopes","sequence":"additional","affiliation":[{"name":"Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, Avenida Ipiranga, 2752\/lab 606, Porto Alegre 90610-000, RS, Brazil"},{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0777-9497","authenticated-orcid":false,"given":"Flavia Nathiely Silveira","family":"Fachel","sequence":"additional","affiliation":[{"name":"Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, Avenida Ipiranga, 2752\/lab 606, Porto Alegre 90610-000, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giselle","family":"Martins","sequence":"additional","affiliation":[{"name":"Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, Avenida Ipiranga, 2752\/lab 606, Porto Alegre 90610-000, RS, Brazil"},{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Siebert","sequence":"additional","affiliation":[{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2820-2814","authenticated-orcid":false,"given":"Willian da Silva","family":"Carniel","sequence":"additional","affiliation":[{"name":"Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, Avenida Ipiranga, 2752\/lab 606, Porto Alegre 90610-000, RS, Brazil"},{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tuane Nerissa Alves","family":"Garcez","sequence":"additional","affiliation":[{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helder Ferreira","family":"Teixeira","sequence":"additional","affiliation":[{"name":"Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, Avenida Ipiranga, 2752\/lab 606, Porto Alegre 90610-000, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4977-6662","authenticated-orcid":false,"given":"Ursula","family":"Matte","sequence":"additional","affiliation":[{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"},{"name":"Graduate Program in Genetics and Molecular Biology, Institute of Biosciences, Universidade Federal do Rio Grande do Sul, Porto Alegre 90010-150, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberto","family":"Giugliani","sequence":"additional","affiliation":[{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"},{"name":"Graduate Program in Genetics and Molecular Biology, Institute of Biosciences, Universidade Federal do Rio Grande do Sul, Porto Alegre 90010-150, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guilherme","family":"Baldo","sequence":"additional","affiliation":[{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"},{"name":"Graduate Program in Genetics and Molecular Biology, Institute of Biosciences, Universidade Federal do Rio Grande do Sul, Porto Alegre 90010-150, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0554-0295","authenticated-orcid":false,"given":"\u00c9dina","family":"Poletto","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Stanford University, Stanford, CA 94305, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4189-0436","authenticated-orcid":false,"given":"Roselena Silvestri","family":"Schuh","sequence":"additional","affiliation":[{"name":"Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul, Avenida Ipiranga, 2752\/lab 606, Porto Alegre 90610-000, RS, Brazil"},{"name":"Cells, Tissues, and Genes Research Group, Experimental Research Center, Hospital de Cl\u00ednicas de Porto Alegre, Porto Alegre 90035-903, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1555","DOI":"10.1021\/acs.accounts.9b00106","article-title":"Delivery Aspects of CRISPR\/Cas for In Vivo Genome Editing","volume":"52","author":"Wilbie","year":"2019","journal-title":"Acc. Chem. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1128\/JB.00580-17","article-title":"History of CRISPR-Cas from encounter with a mysterious repeated sequence to genome editing technology","volume":"200","author":"Ishino","year":"2018","journal-title":"J. Bacteriol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Lundstrom, K. (2023). Viral Vectors in Gene Therapy: Where Do We Stand in 2023?. Viruses, 15.","DOI":"10.3390\/v15030698"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1016\/j.ijpharm.2017.10.030","article-title":"Cationic nanoemulsions as nucleic acids delivery systems","volume":"534","author":"Teixeira","year":"2017","journal-title":"Int. J. Pharm."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1038\/cr.2017.16","article-title":"A non-viral CRISPR\/Cas9 delivery system for therapeutically targeting HBV DNA and pcsk9 in vivo","volume":"27","author":"Jiang","year":"2017","journal-title":"Cell Res."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Glass, Z., Lee, M., Li, Y., and Xu, Q. (2018). Engineering the Delivery System for CRISPR-based Genome Editing. Trends Biotechnol., 36.","DOI":"10.1016\/j.tibtech.2017.11.006"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.jconrel.2018.08.031","article-title":"In vivo genome editing of mucopolysaccharidosis I mice using the CRISPR\/Cas9 system","volume":"288","author":"Schuh","year":"2018","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1038\/s41434-019-0113-4","article-title":"Neonatal nonviral gene editing with the CRISPR\/Cas9 system improves some cardiovascular, respiratory, and bone disease features of the mucopolysaccharidosis I phenotype in mice","volume":"27","author":"Schuh","year":"2020","journal-title":"Gene Ther."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Guo, C., Ma, X., Gao, F., and Guo, Y. (2023). Off-target effects in CRISPR\/Cas9 gene editing. Front. Bioeng. Biotechnol., 11.","DOI":"10.3389\/fbioe.2023.1143157"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e187","DOI":"10.1093\/nar\/gkt772","article-title":"Efficient generation of large-scale genome-modified mice using gRNA and CAS9 endonuclease","volume":"41","author":"Fujii","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e214","DOI":"10.1038\/mtna.2014.64","article-title":"COSMID: A Web-based Tool for Identifying and Validating CRISPR\/Cas off-target Sites","volume":"3","author":"Cradick","year":"2014","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/j.jcis.2018.06.058","article-title":"Physicochemical properties of cationic nanoemulsions and liposomes obtained by microfluidization complexed with a single plasmid or along with an oligonucleotide: Implications for CRISPR\/Cas technology","volume":"530","author":"Schuh","year":"2018","journal-title":"J. Colloid Interface Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1007\/s11095-010-0338-9","article-title":"Hydrodynamic gene delivery and its applications in pharmaceutical research","volume":"28","author":"Bonamassa","year":"2011","journal-title":"Pharm. Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/S0168-3659(00)00293-5","article-title":"Characteristics and biodistribution of cationic liposomes and their DNA complexes","volume":"69","author":"Ishiwata","year":"2000","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1016\/j.ymthe.2005.08.021","article-title":"Mechanistic studies of sequential injection of cationic liposome and plasmid DNA","volume":"13","author":"Zhang","year":"2006","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.addr.2012.09.037","article-title":"Liposomal drug delivery systems: From concept to clinical applications","volume":"65","author":"Allen","year":"2013","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Cataldi, M., Vigliotti, C., Mosca, T., Cammarota, M.R., and Capone, D. (2017). Emerging role of the spleen in the pharmacokinetics of monoclonal antibodies, nanoparticles and exosomes. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18061249"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1038\/sj.gt.3302891","article-title":"Gene therapy progress and prospects: Hydrodynamic gene delivery","volume":"14","author":"Herweijer","year":"2007","journal-title":"Gene Ther."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/S0065-2660(05)54004-5","article-title":"Hydrodynamic Delivery","volume":"54","author":"Knapp","year":"2005","journal-title":"Adv. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1089\/10430340050015734","article-title":"Intravenous Injection of Naked DNA Encoding Secreted flt3 Ligand Dramatically Increases the Number of Dendritic Cells and Natural Killer Cells In Vivo","volume":"11","author":"He","year":"2000","journal-title":"Hum. Gene Ther."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5113","DOI":"10.1158\/1538-7445.AM2018-5113","article-title":"Spontaneous Tumors in Aging Colonies of C57BL\/6J Mice","volume":"78","author":"Schile","year":"2018","journal-title":"Cancer Res."},{"key":"ref_22","unstructured":"(2024, September 29). Some Tumor Data for This Paper were Retrieved from the Mouse Models of Human Cancer Database (MMHCdb), Mouse Genome Informatics, The Jackson Laboratory, Bar Harbor, Maine. World Wide Web. Available online: https:\/\/tumor.informatics.jax.org\/mtbwi\/dynamicGrid.do."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e264","DOI":"10.1038\/mtna.2015.37","article-title":"Off-target Effects in CRISPR\/Cas9-mediated Genome Engineering","volume":"4","author":"Zhang","year":"2015","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7473","DOI":"10.1093\/nar\/gku402","article-title":"CRISPR\/Cas9 systems have off-target activity with insertions or deletions between target DNA and guide RNA sequences","volume":"42","author":"Lin","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1380","DOI":"10.1016\/j.cell.2013.08.021","article-title":"Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity","volume":"154","author":"Ran","year":"2013","journal-title":"Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.biotechadv.2016.12.003","article-title":"Detection of on-target and off-target mutations generated by CRISPR\/Cas9 and other sequence-specific nucleases","volume":"35","author":"Zischewski","year":"2017","journal-title":"Biotechnol. Adv."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1038\/s41596-020-00431-y","article-title":"Tools for experimental and computational analyses of off-target editing by programmable nucleases","volume":"16","author":"Bao","year":"2021","journal-title":"Nat. Protoc."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Liang, D., Gutierrez, N.M., Chen, T., Lee, Y., Park, S.-W., Ma, H., Koski, A., Ahmed, R., Darby, H., and Li, Y. (2020). Frequent Gene Conversion in Human Embryos Induced by Double Strand Breaks, CHS.","DOI":"10.1101\/2020.06.19.162214"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.canlet.2019.01.017","article-title":"CRISPR-Cas9 for cancer therapy: Opportunities and challenges","volume":"447","author":"Chen","year":"2019","journal-title":"Cancer Lett."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Xie, Y., Wang, M., Gu, L., and Wang, Y. (2022). CRISPR\/Cas9-mediated knock-in strategy at the Rosa26 locus in cattle fetal fibroblasts. PLoS ONE, 17.","DOI":"10.1371\/journal.pone.0276811"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1007\/s00109-020-01893-z","article-title":"Mitigating off-target effects in CRISPR\/Cas9-mediated in vivo gene editing","volume":"98","author":"Han","year":"2020","journal-title":"J. Mol. Med."},{"key":"ref_32","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_33","doi-asserted-by":"crossref","unstructured":"Naeem, M., Majeed, S., Hoque, M.Z., and Ahmad, I. (2020). Latest Developed Strategies to Minimize the off-Target Effects in CRISPR-Cas-Mediated Genome Editing. Cells, 9.","DOI":"10.3390\/cells9071608"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Martin, F., S\u00e1nchez-Hern\u00e1ndez, S., Guti\u00e9rrez-Guerrero, A., Pinedo-Gomez, J., and Benabdellah, K. (2016). Biased and unbiased methods for the detection of off-target cleavage by CRISPR\/Cas9: An overview. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17091507"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Atkins, A., Chung, C.H., Allen, A.G., Dampier, W., Gurrola, T.E., Sariyer, I.K., Nonnemacher, M.R., and Wigdahl, B. (2021). Off-Target Analysis in Gene Editing and Applications for Clinical Translation of CRISPR\/Cas9 in HIV-1 Therapy. Front. Genome Ed., 3.","DOI":"10.3389\/fgeed.2021.673022"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Yaish, O., Asif, M., and Orenstein, Y. (2022). A systematic evaluation of data processing and problem formulation of CRISPR off-target site prediction. Brief. Bioinform., 23.","DOI":"10.1093\/bib\/bbac157"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1839","DOI":"10.1016\/j.chembiol.2024.09.006","article-title":"Reducing CRISPR-Cas9 off-target effects by optically controlled chemical modifications of guide RNA","volume":"31","author":"Qi","year":"2024","journal-title":"Cell Chem. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1038\/nbt.3117","article-title":"GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases","volume":"33","author":"Tsai","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3596","DOI":"10.1038\/s41467-020-17418-8","article-title":"Prediction-based highly sensitive CRISPR off-target validation using target-specific DNA enrichment","volume":"11","author":"Kang","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1038\/s41467-023-35886-6","article-title":"Assessing and advancing the safety of CRISPR-Cas tools: From DNA to RNA editing","volume":"14","author":"Tao","year":"2023","journal-title":"Nat. Commun."}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/17\/5\/656\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:34:19Z","timestamp":1760031259000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/17\/5\/656"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,17]]},"references-count":40,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2025,5]]}},"alternative-id":["pharmaceutics17050656"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics17050656","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,17]]}}}