{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T14:56:18Z","timestamp":1790693778049,"version":"4.1.0"},"reference-count":62,"publisher":"American Chemical Society (ACS)","issue":"7","license":[{"start":{"date-parts":[[2020,6,23]],"date-time":"2020-06-23T00:00:00Z","timestamp":1592870400000},"content-version":"vor","delay-in-days":18,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/100010663","name":"H2020 European Research Council","doi-asserted-by":"publisher","award":["670216"],"award-info":[{"award-number":["670216"]}],"id":[{"id":"10.13039\/100010663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010668","name":"H2020 Leadership in Enabling and Industrial Technologies","doi-asserted-by":"publisher","award":["634942"],"award-info":[{"award-number":["634942"]}],"id":[{"id":"10.13039\/100010668","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100014440","name":"Ministerio de Ciencia, Innovaci\u00f3n y Universidades","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/100014440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002809","name":"Generalitat de Catalunya","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/501100002809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["CP16\/00094"],"award-info":[{"award-number":["CP16\/00094"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008530","name":"European Regional Development Fund","doi-asserted-by":"publisher","award":["CP16\/00094"],"award-info":[{"award-number":["CP16\/00094"]}],"id":[{"id":"10.13039\/501100008530","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,7,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Mycoplasma species share a set of features, such as lack of a cell wall, streamlined genomes, simplified metabolism, and the use of a deviant genetic code, that make them attractive approximations of what a chassis strain should ideally be. Among them, Mycoplasma pneumoniae arises as a candidate for synthetic biology projects, as it is one of the most deeply characterized bacteria. However, the historical paucity of tools for editing Mycoplasma genomes has precluded the establishment of M. pneumoniae as a suitable chassis strain. Here, we developed an oligonucleotide recombineering method for this strain based on GP35, a ssDNA recombinase originally encoded by a Bacillus subtilis-associated phage. GP35-mediated oligo recombineering is able to carry out point mutations in the M. pneumoniae genome with an efficiency as high as 2.7 \u00d7 10\u20132, outperforming oligo recombineering protocols developed for other bacteria. Gene deletions of different sizes showed a decreasing power trend between efficiency and the scale of the attempted edition. However, the editing rates for all modifications increased when CRISPR\/Cas9 was used to counterselect nonedited cells. This allowed edited clones carrying chromosomal deletions of up to 1.8 kb to be recovered with little to no screening of survivor cells. We envision this technology as a major step toward the use of M. pneumoniae, and possibly other Mycoplasmas, as synthetic biology chassis strains.<\/jats:p>","DOI":"10.1021\/acssynbio.0c00022","type":"journal-article","created":{"date-parts":[[2020,6,5]],"date-time":"2020-06-05T18:00:20Z","timestamp":1591380020000},"page":"1693-1704","source":"Crossref","is-referenced-by-count":39,"title":["<i>Mycoplasma pneumoniae<\/i>\n                    Genome Editing\nBased on Oligo Recombineering and Cas9-Mediated Counterselection"],"prefix":"10.1021","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5263-3702","authenticated-orcid":true,"given":"Carlos","family":"Pin\u0303ero-Lambea","sequence":"first","affiliation":[{"name":"Centre for Genomic Regulation (CRG) , , Dr. Aiguader 88 , ,","place":["Barcelona, Spain, 08003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Garcia-Ramallo","sequence":"additional","affiliation":[{"name":"Centre for Genomic Regulation (CRG) , , Dr. Aiguader 88 , ,","place":["Barcelona, Spain, 08003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sira","family":"Martinez","sequence":"additional","affiliation":[{"name":"Centre for Genomic Regulation (CRG) , , Dr. Aiguader 88 , ,","place":["Barcelona, Spain, 08003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Javier","family":"Delgado","sequence":"additional","affiliation":[{"name":"Centre for Genomic Regulation (CRG) , , Dr. Aiguader 88 , ,","place":["Barcelona, Spain, 08003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis","family":"Serrano","sequence":"additional","affiliation":[{"name":"Centre for Genomic Regulation (CRG) , , Dr. Aiguader 88 , ,","place":["Barcelona, Spain, 08003"]},{"name":"ICREA , Pg. Lluis Companys 23 , ,","place":["Barcelona, Spain, 08010"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Lluch-Senar","sequence":"additional","affiliation":[{"name":"Centre for Genomic Regulation (CRG) , , Dr. Aiguader 88 , ,","place":["Barcelona, Spain, 08003"]},{"name":"Universitat Pompeu Fabra (UPF) , ,","place":["Barcelona, Spain, 08003"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,6,5]]},"reference":[{"issue":"14","key":"2026090309472304600_cit1","doi-asserted-by":"publisher","first-page":"5053","DOI":"10.1074\/jbc.TM117.000368","article-title":"Synthetic\nBiology Strategies for\nImproving Microbial Synthesis of \u201cGreen\u201d Biopolymers","volume":"293","author":"Anderson","year":"2018","journal-title":"J. Biol. Chem."},{"key":"2026090309472304600_cit2","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1186\/s12934-014-0173-5","article-title":"Development of Bio-Based\nFine Chemical Production through Synthetic\nBioengineering","volume":"13","author":"Hara","year":"2014","journal-title":"Microb. Cell Fact."},{"issue":"7","key":"2026090309472304600_cit3","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1016\/j.biotechadv.2017.08.001","article-title":"Bioremediation\n3.0: Engineering Pollutant-Removing Bacteria in the Times of Systemic\nBiology","volume":"35","author":"Dvor\u030ca\u0301k","year":"2017","journal-title":"Biotechnol. Adv."},{"key":"2026090309472304600_cit4","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1016\/j.copbio.2015.05.004","article-title":"Engineered Bacteria as Therapeutic\nAgents","volume":"35","author":"Pin\u0303ero-Lambea","year":"2015","journal-title":"Curr. Opin. Biotechnol."},{"key":"2026090309472304600_cit5","doi-asserted-by":"publisher","first-page":"158","DOI":"10.1038\/msb4100202","article-title":"Synthetic\nBiology: Promises and Challenges","volume":"3","author":"Serrano","year":"2007","journal-title":"Mol. Syst.\nBiol."},{"issue":"6211","key":"2026090309472304600_cit6","doi-asserted-by":"publisher","first-page":"1256272","DOI":"10.1126\/science.1256272","article-title":"Synthetic Biology.\nGenomically Encoded Analog Memory with Precise\nin Vivo DNA Writing in Living Cell Populations","volume":"346","author":"Farzadfard","year":"2014","journal-title":"Science"},{"issue":"5931","key":"2026090309472304600_cit7","doi-asserted-by":"publisher","first-page":"1199","DOI":"10.1126\/science.1172005","article-title":"Synthetic Gene Networks That Count","volume":"324","author":"Friedland","year":"2009","journal-title":"Science"},{"issue":"6767","key":"2026090309472304600_cit8","doi-asserted-by":"publisher","first-page":"339","DOI":"10.1038\/35002131","article-title":"Construction\nof a Genetic Toggle\nSwitch in Escherichia Coli","volume":"403","author":"Gardner","year":"2000","journal-title":"Nature"},{"issue":"4","key":"2026090309472304600_cit9","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1021\/sb500252a","article-title":"Programming Controlled Adhesion of E. Coli to Target Surfaces, Cells,\nand Tumors with Synthetic Adhesins","volume":"4","author":"Pin\u0303ero-Lambea","year":"2015","journal-title":"ACS Synth.\nBiol."},{"key":"2026090309472304600_cit10","doi-asserted-by":"publisher","first-page":"521","DOI":"10.1038\/msb.2011.55","article-title":"Engineering Microbes\nto Sense and Eradicate Pseudomonas Aeruginosa, a Human Pathogen","volume":"7","author":"Saeidi","year":"2011","journal-title":"Mol. Syst. Biol."},{"issue":"9","key":"2026090309472304600_cit11","doi-asserted-by":"publisher","first-page":"857","DOI":"10.1038\/nbt.4222","article-title":"Development of a Synthetic Live Bacterial Therapeutic for the Human\nMetabolic Disease Phenylketonuria","volume":"36","author":"Isabella","year":"2018","journal-title":"Nat. Biotechnol."},{"issue":"475","key":"2026090309472304600_cit12","doi-asserted-by":"publisher","first-page":"eaau7975","DOI":"10.1126\/scitranslmed.aau7975","article-title":"An Engineered\nE.\nColi Nissle Improves Hyperammonemia and Survival in Mice and Shows\nDose-Dependent Exposure in Healthy Humans","volume":"11","author":"Kurtz","year":"2019","journal-title":"Sci.\nTransl. Med."},{"issue":"12","key":"2026090309472304600_cit13","doi-asserted-by":"publisher","first-page":"1328","DOI":"10.1021\/acssynbio.6b00256","article-title":"The Next\nGeneration of Synthetic Biology Chassis: Moving Synthetic Biology\nfrom the Laboratory to the Field","volume":"5","author":"Adams","year":"2016","journal-title":"ACS Synth.\nBiol."},{"issue":"4","key":"2026090309472304600_cit14","doi-asserted-by":"publisher","first-page":"1094","DOI":"10.1128\/MMBR.62.4.1094-1156.1998","article-title":"Molecular Biology and\nPathogenicity\nof Mycoplasmas","volume":"62","author":"Razin","year":"1998","journal-title":"Microbiol. Mol. Biol. Rev."},{"issue":"5957","key":"2026090309472304600_cit15","doi-asserted-by":"publisher","first-page":"1268","DOI":"10.1126\/science.1176951","article-title":"Transcriptome Complexity\nin a Genome-Reduced Bacterium","volume":"326","author":"Gu\u0308ell","year":"2009","journal-title":"Science"},{"issue":"5957","key":"2026090309472304600_cit16","doi-asserted-by":"publisher","first-page":"1263","DOI":"10.1126\/science.1177263","article-title":"Impact of\nGenome Reduction\non Bacterial Metabolism and Its Regulation","volume":"326","author":"Yus","year":"2009","journal-title":"Science"},{"issue":"1","key":"2026090309472304600_cit17","doi-asserted-by":"publisher","first-page":"511","DOI":"10.1038\/msb.2011.38","article-title":"Quantification of MRNA\nand Protein and Integration with Protein Turnover in a Bacterium","volume":"7","author":"Maier","year":"2011","journal-title":"Mol. Syst. Biol."},{"key":"2026090309472304600_cit18","doi-asserted-by":"publisher","first-page":"585","DOI":"10.1038\/msb.2012.16","article-title":"Transcription\nStart Site Associated RNAs in Bacteria","volume":"8","author":"Yus","year":"2012","journal-title":"Mol.\nSyst. Biol."},{"issue":"1","key":"2026090309472304600_cit19","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1003191","article-title":"Comprehensive\nMethylome Characterization\nof Mycoplasma Genitalium and Mycoplasma Pneumoniae at Single-Base\nResolution","volume":"9","author":"Lluch-Senar","year":"2013","journal-title":"PLoS Genet."},{"issue":"5","key":"2026090309472304600_cit20","doi-asserted-by":"publisher","first-page":"4030","DOI":"10.3892\/mmr.2016.5765","article-title":"Insights into\nthe Pathogenesis of\nMycoplasma Pneumoniae","volume":"14","author":"He","year":"2016","journal-title":"Mol. Med. Rep."},{"issue":"9","key":"2026090309472304600_cit21","doi-asserted-by":"publisher","first-page":"5227","DOI":"10.1073\/pnas.96.9.5227","article-title":"Disruption of Gene Mg218 of Mycoplasma\nGenitalium through Homologous Recombination Leads to an Adherence-Deficient\nPhenotype","volume":"96","author":"Dhandayuthapani","year":"1999","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"issue":"19","key":"2026090309472304600_cit22","doi-asserted-by":"publisher","first-page":"5645","DOI":"10.1128\/JB.183.19.5645-5650.2001","article-title":"Peptide Methionine Sulfoxide Reductase (MsrA) Is a\nVirulence Determinant in Mycoplasma Genitalium","volume":"183","author":"Dhandayuthapani","year":"2001","journal-title":"J. Bacteriol."},{"issue":"4","key":"2026090309472304600_cit23","doi-asserted-by":"publisher","first-page":"1029","DOI":"10.1111\/j.1365-2958.2008.06343.x","article-title":"Deletion of\nthe Mycoplasma Genitalium MG_217 Gene Modifies Cell Gliding Behaviour\nby Altering Terminal Organelle Curvature","volume":"69","author":"Burgos","year":"2008","journal-title":"Mol.\nMicrobiol."},{"issue":"20","key":"2026090309472304600_cit24","doi-asserted-by":"publisher","first-page":"6951","DOI":"10.1128\/AEM.00056-10","article-title":"Enhancement\nof Targeted Homologous Recombination in Mycoplasma Mycoides Subsp.\nCapri by Inclusion of Heterologous RecA","volume":"76","author":"Allam","year":"2010","journal-title":"Appl.\nEnviron. Microbiol."},{"issue":"15","key":"2026090309472304600_cit25","doi-asserted-by":"publisher","first-page":"5297","DOI":"10.1128\/AEM.00024-10","article-title":"Targeted\nChromosomal Knockouts in Mycoplasma Pneumoniae","volume":"76","author":"Krishnakumar","year":"2010","journal-title":"Appl. Environ. Microbiol."},{"issue":"5","key":"2026090309472304600_cit26","doi-asserted-by":"publisher","first-page":"1261","DOI":"10.1111\/j.1365-2958.2010.07288.x","article-title":"The Mycoplasma Genitalium\nMG352-Encoded Protein Is a Holliday Junction Resolvase That Has a\nNon-Functional Orthologue in Mycoplasma Pneumoniae","volume":"77","author":"Sluijter","year":"2010","journal-title":"Mol. Microbiol."},{"issue":"5","key":"2026090309472304600_cit27","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0038301","article-title":"The RuvA Homologues from Mycoplasma Genitalium and\nMycoplasma Pneumoniae Exhibit Unique Functional Characteristics","volume":"7","author":"Sluijter","year":"2012","journal-title":"PLoS One"},{"issue":"10","key":"2026090309472304600_cit28","doi-asserted-by":"publisher","first-page":"4923","DOI":"10.1093\/nar\/gkv422","article-title":"A Novel Sigma Factor Reveals a Unique Regulon Controlling\nCell-Specific Recombination in Mycoplasma Genitalium","volume":"43","author":"Torres-Puig","year":"2015","journal-title":"Nucleic Acids Res."},{"issue":"1","key":"2026090309472304600_cit29","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.ijmm.2006.11.001","article-title":"Tools for the Genetic\nAnalysis of Mycoplasma","volume":"297","author":"Halbedel","year":"2007","journal-title":"Int. J. Med. Microbiol."},{"issue":"5867","key":"2026090309472304600_cit30","doi-asserted-by":"publisher","first-page":"1215","DOI":"10.1126\/science.1151721","article-title":"Complete Chemical Synthesis, Assembly, and Cloning\nof a Mycoplasma Genitalium Genome","volume":"319","author":"Gibson","year":"2008","journal-title":"Science"},{"issue":"5987","key":"2026090309472304600_cit31","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1126\/science.1190719","article-title":"Creation of a Bacterial\nCell Controlled by a Chemically Synthesized Genome","volume":"329","author":"Gibson","year":"2010","journal-title":"Science"},{"issue":"8","key":"2026090309472304600_cit32","doi-asserted-by":"publisher","first-page":"2558","DOI":"10.1093\/nar\/gkq119","article-title":"Cloning\nWhole Bacterial Genomes in\nYeast","volume":"38","author":"Benders","year":"2010","journal-title":"Nucleic Acids Res."},{"issue":"8","key":"2026090309472304600_cit33","doi-asserted-by":"publisher","first-page":"2570","DOI":"10.1093\/nar\/gkq099","article-title":"Tandem Repeat Coupled with Endonuclease\nCleavage (TREC): A Seamless Modification Tool for Genome Engineering\nin Yeast","volume":"38","author":"Noskov","year":"2010","journal-title":"Nucleic Acids Res."},{"key":"2026090309472304600_cit34","doi-asserted-by":"publisher","first-page":"1180","DOI":"10.1186\/1471-2164-15-1180","article-title":"TREC-IN:\nGene Knock-in Genetic Tool for Genomes Cloned in Yeast","volume":"15","author":"Chandran","year":"2014","journal-title":"BMC Genomics"},{"issue":"11","key":"2026090309472304600_cit35","doi-asserted-by":"publisher","first-page":"2547","DOI":"10.1021\/acssynbio.9b00224","article-title":"CReasPy-Cloning:\nA Method for Simultaneous Cloning and Engineering of Megabase-Sized\nGenomes in Yeast Using the CRISPR-Cas9 System","volume":"8","author":"Ruiz","year":"2019","journal-title":"ACS Synth. Biol."},{"issue":"5838","key":"2026090309472304600_cit36","doi-asserted-by":"publisher","first-page":"632","DOI":"10.1126\/science.1144622","article-title":"Genome Transplantation\nin Bacteria: Changing One Species to Another","volume":"317","author":"Lartigue","year":"2007","journal-title":"Science"},{"key":"2026090309472304600_cit37","doi-asserted-by":"publisher","first-page":"664","DOI":"10.3389\/fmicb.2019.00664","article-title":"Removal of a Subset of Non-Essential Genes Fully Attenuates\na Highly Virulent Mycoplasma Strain","volume":"10","author":"Jores","year":"2019","journal-title":"Front.\nMicrobiol."},{"issue":"17","key":"2026090309472304600_cit38","doi-asserted-by":"publisher","first-page":"8501","DOI":"10.1093\/nar\/gkw688","article-title":"Impact of\nDonor-Recipient Phylogenetic Distance on Bacterial Genome Transplantation","volume":"44","author":"Labroussaa","year":"2016","journal-title":"Nucleic Acids Res."},{"issue":"12","key":"2026090309472304600_cit39","doi-asserted-by":"publisher","first-page":"5151","DOI":"10.1007\/s00253-015-6485-5","article-title":"A High-Efficiency\nRecombineering\nSystem with PCR-Based SsDNA in Bacillus Subtilis Mediated by the Native\nPhage Recombinase GP35","volume":"99","author":"Sun","year":"2015","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"2026090309472304600_cit40","doi-asserted-by":"publisher","first-page":"1.16.1","DOI":"10.1002\/0471142727.mb0116s106","article-title":"Recombineering: Genetic Engineering in Bacteria Using\nHomologous Recombination","volume":"106","author":"Thomason","year":"2014","journal-title":"Curr. Protoc. Mol.\nBiol."},{"issue":"12","key":"2026090309472304600_cit41","doi-asserted-by":"publisher","first-page":"6742","DOI":"10.1073\/pnas.121164898","article-title":"High Efficiency Mutagenesis, Repair,\nand Engineering of Chromosomal DNA Using Single-Stranded Oligonucleotides","volume":"98","author":"Ellis","year":"2001","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"issue":"5","key":"2026090309472304600_cit42","doi-asserted-by":"publisher","first-page":"1626","DOI":"10.1073\/pnas.0709089105","article-title":"Identification and\nAnalysis of Recombineering Functions from Gram-Negative and Gram-Positive\nBacteria and Their Phages","volume":"105","author":"Datta","year":"2008","journal-title":"Proc. Natl. Acad.\nSci. U. S. A."},{"issue":"7257","key":"2026090309472304600_cit43","doi-asserted-by":"publisher","first-page":"894","DOI":"10.1038\/nature08187","article-title":"Programming\nCells by Multiplex Genome\nEngineering and Accelerated Evolution","volume":"460","author":"Wang","year":"2009","journal-title":"Nature"},{"issue":"1","key":"2026090309472304600_cit44","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1016\/j.mimet.2005.02.011","article-title":"An Improved Arbitrary\nPrimed PCR Method for Rapid Characterization of Transposon Insertion\nSites","volume":"63","author":"Das","year":"2005","journal-title":"J. Microbiol. Methods"},{"issue":"1","key":"2026090309472304600_cit45","doi-asserted-by":"publisher","first-page":"176","DOI":"10.1111\/1751-7915.12846","article-title":"A Standardized Workflow for Surveying\nRecombinases Expands Bacterial Genome-Editing Capabilities","volume":"11","author":"Ricaurte","year":"2018","journal-title":"Microb. Biotechnol."},{"issue":"1","key":"2026090309472304600_cit46","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1128\/mBio.00067-18","article-title":"Efficient and Scalable Precision\nGenome Editing in\nStaphylococcus Aureus through Conditional Recombineering and CRISPR\/Cas9-Mediated\nCounterselection","volume":"9","author":"Penewit","year":"2018","journal-title":"mBio"},{"issue":"1","key":"2026090309472304600_cit47","doi-asserted-by":"publisher","first-page":"0","DOI":"10.2225\/vol11-issue1-fulltext-1","article-title":"Multiple Pulses Improve Electroporation Efficiency\nin Agrobacterium Tumefaciens","volume":"11","author":"Mahmood","year":"2008","journal-title":"Electron. J. Biotechnol."},{"issue":"6213","key":"2026090309472304600_cit48","doi-asserted-by":"publisher","first-page":"1258096","DOI":"10.1126\/science.1258096","article-title":"Genome Editing. The New Frontier of Genome Engineering\nwith CRISPR-Cas9","volume":"346","author":"Doudna","year":"2014","journal-title":"Science"},{"issue":"3","key":"2026090309472304600_cit49","doi-asserted-by":"publisher","first-page":"233","DOI":"10.1038\/nbt.2508","article-title":"RNA-Guided\nEditing of Bacterial Genomes Using CRISPR-Cas Systems","volume":"31","author":"Jiang","year":"2013","journal-title":"Nat. Biotechnol."},{"issue":"9","key":"2026090309472304600_cit50","doi-asserted-by":"publisher","first-page":"4243","DOI":"10.1093\/nar\/gkw223","article-title":"Consequences of Cas9\nCleavage in the Chromosome of\nEscherichia Coli","volume":"44","author":"Cui","year":"2016","journal-title":"Nucleic Acids Res."},{"issue":"5","key":"2026090309472304600_cit51","doi-asserted-by":"publisher","first-page":"1076","DOI":"10.1111\/1751-7915.13453","article-title":"CRISPR\/Cas9-Enhanced\nSsDNA Recombineering for Pseudomonas Putida","volume":"12","author":"Aparicio","year":"2019","journal-title":"Microb. Biotechnol."},{"key":"2026090309472304600_cit52","doi-asserted-by":"publisher","first-page":"15096","DOI":"10.1038\/srep15096","article-title":"The No-SCAR (Scarless\nCas9 Assisted Recombineering)\nSystem for Genome Editing in Escherichia Coli","volume":"5","author":"Reisch","year":"2015","journal-title":"Sci. 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