ArticleMicrobial cell factories2019
Optimizing a CRISPR-Cpf1-based genome engineering system for Corynebacterium glutamicum.
Article in Microbial cell factories, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- Tailoring Corynebacterium glutamicum for Sustainable Biomanufacturing: From Traditional to Cutting-Edge Technologies.Molecular biotechnology · 2026Review
- Advances in gene editing tools for four typical Gram-positive bacteria.Frontiers in microbiology · 2026Review
- Development of a CRISPR/Cas9 genome editing toolbox forSynthetic and systems biotechnology · 2025Article
- Expanding the CRISPR Toolbox for Engineering Lycopene Biosynthesis inMicroorganisms · 2024Article
- Establishment of CRISPR-Cpf1-assisted gene editing tool and engineering of 4-hydroxyisoleucine biosynthesis in Corynebacterium glutamicum.World journal of microbiology & biotechnology · 2023Article
- Single and multiplexed gene repression in solventogenicSynthetic and systems biotechnology · 2023Article
- Review
- Enhanced production of D-pantothenic acid in Corynebacterium glutamicum using an efficient CRISPR-Cpf1 genome editing method.Microbial cell factories · 2023Article
- Synthetic biology tools for engineeringComputational and structural biotechnology journal · 2023Review
- Development of an in vivo cleavable donor plasmid for targeted transgene integration by CRISPR-Cas9 and CRISPR-Cas12a.Scientific reports · 2022Article
- CRISPR-assisted rational flux-tuning and arrayed CRISPRi screening of an L-proline exporter for L-proline hyperproduction.Nature communications · 2022Article
- Bacterial genome reductions: Tools, applications, and challenges.Frontiers in genome editing · 2022Review
- CRISPRi-Library-Guided Target Identification for Engineering Carotenoid Production byMicroorganisms · 2021Article
- Advances and Perspectives for Genome Editing Tools ofFrontiers in microbiology · 2021Review
- Single-Base Genome Editing inJournal of microbiology and biotechnology · 2020Article
- Multiplex gene editing and large DNA fragment deletion by the CRISPR/Cpf1-RecE/T system in Corynebacterium glutamicum.Journal of industrial microbiology & biotechnology · 2020Article
- Obtaining a series of native gradient promoter-5'-UTR sequences in Corynebacterium glutamicum ATCC 13032.Microbial cell factories · 2020Article
- The Versatile Type V CRISPR Effectors and Their Application Prospects.Frontiers in cell and developmental biology · 2020Review
- Barriers to genome editing with CRISPR in bacteria.Journal of industrial microbiology & biotechnology · 2019Review
- Recent Advances of L-ornithine Biosynthesis in Metabolically EngineeredFrontiers in bioengineering and biotechnology · 2019Review
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5 authors.
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Abstract
backgroundCorynebacterium glutamicum is an important industrial strain for the production of a diverse range of chemicals. Cpf1 nucleases are highly specific and programmable, with efficiencies comparable to those of Cas9. Although the Francisella novicida (Fn) CRISPR-Cpf1 system has been adapted for genome editing in C. glutamicum, the editing efficiency is currently less than 15%, due to false positives caused by the poor targeting efficiency of the crRNA.
resultsTo address this limitation, a screening strategy was developed in this study to systematically evaluate crRNA targeting efficiency in C. glutamicum. We quantitatively examined various parameters of the C. glutamicum CRISPR-Cpf1 system, including the protospacer adjacent motif (PAM) sequence, the length of the spacer sequence, and the type of repair template. We found that the most efficient C. glutamicum crRNA contained a 5'-NYTV-3' PAM and a 21 bp spacer sequence. Moreover, we observed that linear DNA could be used to repair double strand breaks.
conclusionsHere, we identified optimized PAM-related parameters for the CRISPR-Cpf1 system in C. glutamicum. Our study sheds light on the function of the FnCpf1 endonuclease and Cpf1-based genome editing. This optimized system, with higher editing efficiency, could be used to increase the production of bulk chemicals, such as isobutyrate, in C. glutamicum.
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