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Efficient genome editing for Pseudomonas aeruginosa using CRISPR-Cas12a
- Source :
- Gene. 790:145693
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- The CRISPR-Cas12a system has been demonstrated as an attractive tool for bacterial genome engineering. In particular, FnCas12a recognizes protospacer-adjacent motif (PAM) sites with medium or low GC content, which complements the Cas9-based systems. Here we explored Francisella novicida Cas12a (FnCas12a) for genome editing in Pseudomonas aeruginosa. By using a two-plasmid system expressing the constitutive FnCas12a nuclease, the inducible λRed recombinase, a CRISPR RNA (crRNA), we achieved gene deletion, insertion and replacement with high efficiency (in most cases > 75%), including the deletion of large DNA fragments up to 15 kb and the serial deletion of duplicate gene clusters. This work should provide a useful and complementary addition to the genome engineering toolbox for the study of P. aeruginosa biology and physiology.
- Subjects :
- Gene Editing
0301 basic medicine
Trans-activating crRNA
Cas9
General Medicine
Bacterial genome size
Computational biology
Biology
Genome engineering
03 medical and health sciences
030104 developmental biology
0302 clinical medicine
Bacterial Proteins
Genome editing
030220 oncology & carcinogenesis
Pseudomonas aeruginosa
Genetics
Recombinase
CRISPR
CRISPR-Cas Systems
Gene
Genome, Bacterial
Plasmids
Subjects
Details
- ISSN :
- 03781119
- Volume :
- 790
- Database :
- OpenAIRE
- Journal :
- Gene
- Accession number :
- edsair.doi.dedup.....f1a9ad243a141c3cb0f327d4eabc4012