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- Title
Self-cloning CRISPR/Cpf1 facilitated genome editing in Saccharomyces cerevisiae.
- Authors
Li, Zhen-Hai; Wang, Feng-Qing; Wei, Dong-Zhi
- Abstract
Background: Saccharomyces cerevisiae is one of the most important industrial microorganisms. A robust genome editing tool is vital for both fundamental research and applications. To save the time and labor consumed in the procedure of genome editing, a self-cloning CRISPR/Cpf1 system (scCRISPR/Cpf1), in which a self-cleaving plasmid and PCR-generated site-specific crRNA fragment were included, was developed.Results: Using scCRISPR/Cpf1 as the genetic tool, simple and fast singleplex and multiplex genomic integration of in vivo assembled DNA parts were investigated. Moreover, we validate the applicability of scCRISPR/Cpf1 for cell factory development by creating a patchoulol production strain through two rounds of iterative genomic integration. The results showed that scCRISPR/Cpf1 enables singleplex and tripleplex genomic integration of in vivo assembled DNA parts with efficiencies of 80 and 32%, respectively. Furthermore, the patchoulol production strain was successfully and rapidly engineered and optimized through two rounds of iterative genomic integration by scCRISPR/Cpf1.Conclusions: scCRISPR/Cpf1 allows for CRISPR/Cpf1-facilitated genome editing by circumventing the step to clone a site-specific crRNA plasmid without compromising efficiency in S. cerevisiae. This method enriches the current set of tools available for strain engineering in S. cerevisiae.<graphic></graphic>
- Subjects
CRISPRS; GENOME editing; FUNGAL genetics; PLASMID genetics; SACCHAROMYCES cerevisiae; CLONING
- Publication
Bioresources & Bioprocessing, 2018, Vol 5, Issue 1, p1
- ISSN
2197-4365
- Publication type
Article
- DOI
10.1186/s40643-018-0222-8