Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas12a (Cpf1) has emerged as an effective genome editing tool in many organisms. Here, we developed and optimized a CRISPR-Cas12a-assisted recombineering system to facilitate genetic manipulation in bacteria. Using this system, point mutations, deletions, insertions, and gene replacements can be easily generated on the chromosome or native plasmids in Escherichia coli, Yersinia pestis, and Mycobacterium smegmatis. Because CRISPR-Cas12a-assisted recombineering does not require introduction of an antibiotic resistance gene into the chromosome to select for recombinants, it is an efficient approach for generating markerless and scarless mutations in bacteria.
CITATION STYLE
Yan, M. Y., Yan, H. Q., Ren, G. X., Zhao, J. P., Guo, X. P., & Sun, Y. C. (2017). CRISPR-Cas12a-assisted recombineering in bacteria. Applied and Environmental Microbiology, 83(17). https://doi.org/10.1128/AEM.00947-17
Mendeley helps you to discover research relevant for your work.