Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomonas using phage-encoded homologous recombination
Engineering Microbiology(2022)
摘要
Phage-encoded homologous recombination (PEHR) is an efficient tool for bacterial genome editing. We previously developed and utilized a Pseudomonas-specific PEHR system. However, when using the PEHR system for Pseudomonas genome editing, false positives can be a problem. In this study, we combined a compact Cascade-Cas3 system from P. aeruginosa (PaeCas3c) with a Pseudomonas-specific PEHR system, and the results of our recombineering assay showed that this compact Cascade-Cas3 system can significantly improve PEHR recombineering accuracy.
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关键词
Phage-encoded homologous recombination,Pseudomonas,Cascade-Cas3 system,Genome engineering
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