A single-plasmid-based, easily curable CRISPR/Cas9 system for rapid, iterative genome editing in Pseudomonas putida KT2440
We developed and applied a rapid all-in-one plasmid CRISPR/Cas9 system for genome editing in P. putida. This system requires less than 1.5 days for one edit due to simplified plasmid construction, electroporation and curing processes, thus accelerating the cycle of genome editing. To our knowledge, this is the fastest iterative genome editing system for P. putida. Using this system, we rapidly engineered P. putida for valencene biosynthesis for the first time, showcasing the system's potential for expanding biotechnological applications.
Authors
- Jianmin Xing
- Qifeng Wen
- Ida Putu Wiweka Dharmasiddhi
- Jin Li (ORCID: https://orcid.org/0000-0001-9201-2321)
- Jinjin Chen (ORCID: https://orcid.org/0000-0002-3386-5321)
- Yilan Liu (ORCID: https://orcid.org/0000-0002-0923-1425)
- Maohua Yang (ORCID: https://orcid.org/0000-0002-4410-0949)
Institutions
- University of Waterloo (CA)
- Institute of Process Engineering (CN)
- University of Chinese Academy of Sciences (CN)
Publication Details
- Journal
- Microbial Cell Factories
- Published
- 2024-12-29
- DOI
- https://doi.org/10.1186/s12934-024-02634-4
- Citations
- 9
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 2.10
Funders
- University of Waterloo
- Natural Sciences and Engineering Research Council of Canada
- National Key Research and Development Program of China