The DOI matches its Crossref record; 35 of the 35 references it cites were found in a registry or an index.
CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum
Open accessAbstract
Corynebacterium glutamicum is an important industrial metabolite producer that is difficult to genetically engineer. Although the Streptococcus pyogenes (Sp) CRISPR-Cas9 system has been adapted for genome editing of multiple bacteria, it cannot be introduced into C. glutamicum. Here we report a Francisella novicida (Fn) CRISPR-Cpf1-based genome-editing method for C. glutamicum. CRISPR-Cpf1, combined with single-stranded DNA (ssDNA) recombineering, precisely introduces small changes into the bacterial genome at efficiencies of 86–100%. Large gene deletions and insertions are also obtained using an all-in-one plasmid consisting of Fn Cpf1, CRISPR RNA, and homologous arms. The two CRISPR-Cpf1-assisted systems enable N iterative rounds of genome editing in 3 N+4 or 3 N+2 days. A proof-of-concept, codon saturation mutagenesis at G149 of γ-glutamyl kinase relieves L-proline inhibition using Cpf1-assisted ssDNA recombineering. Thus, CRISPR-Cpf1-based genome editing provides a highly efficient tool for genetic engineering of Corynebacterium and other bacteria that cannot utilize the Sp CRISPR-Cas9 system.
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Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System
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Cited by
Cas9 versus Cas12a/Cpf1: Structure–function comparisons and implications for genome editing
Swarts, D. C., & Jinek, M. (2018) · WIREs RNA · doi:10.1002/wrna.1481 · Cited by 267
Application of different types of CRISPR/Cas-based systems in bacteria
Liu, Z., Dong, H., Cui, Y., Cong, L., & Zhang, D. (2020) · Microbial Cell Factories · doi:10.1186/s12934-020-01431-z · Cited by 185
Cas9 off-target binding to the promoter of bacterial genes leads to silencing and toxicity
Rostain, W., Grebert, T., Vyhovskyi, D., Pizarro, P. T., Tshinsele-Van Bellingen, G., Cui, L., & Bikard, D. (2023) · Nucleic Acids Research · doi:10.1093/nar/gkad170 · Cited by 85
Current advances in CRISPR-Cas-mediated gene editing and regulation in cyanobacteria
Dong, Z., Chen, L., Wang, Y., Sun, T., & Zhang, W. (2024) · Blue Biotechnology · doi:10.1186/s44315-024-00009-3 · Cited by 16
Metadata
- Authors
- Jiang, Y., Qian, F., Yang, J., Liu, Y., Dong, F., Xu, C., Sun, B., Chen, B., Xu, X., Li, Y., Wang, R., & Yang, S.
- Venue
- Nature Communications
- Published
- 2017-05-04
- Volume
- 8
- Issue
- 1
- Pages
- 15179-15179
- ISSN
- 2041-1723 (ISSN-L)
- OpenAlex
- W2610391087
- Cited by
- 389
- References
- 35
- FWCI
- 14.66
- Type
- journal article
- Language
- en
- Open access
- gold
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- Jiang, Y., Qian, F., Yang, J., Liu, Y., Dong, F., Xu, C., Sun, B., Chen, B., Xu, X., Li, Y., Wang, R., & Yang, S. (2017). CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum. Nature Communications. https://doi.org/10.1038/ncomms15179
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