ArticleJournal of industrial microbiology & biotechnology2020
Multiplex gene editing and large DNA fragment deletion by the CRISPR/Cpf1-RecE/T system in Corynebacterium glutamicum.
Article in Journal of industrial microbiology & biotechnology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- Tailoring Corynebacterium glutamicum for Sustainable Biomanufacturing: From Traditional to Cutting-Edge Technologies.Molecular biotechnology · 2026Review
- Biotechnological Approaches to Probiotics and Postbiotics through the Gut-Brain Axis Modulation.Iranian biomedical journal · 2026Article
- Advances in gene editing tools for four typical Gram-positive bacteria.Frontiers in microbiology · 2026Review
- Being a better version of yourself: genetically engineered probiotic bacteria as host defense enhancers in the control of intestinal pathogens.Gut microbes · 2025Review
- Development of a CRISPR/Cas9 genome editing toolbox forSynthetic and systems biotechnology · 2025Article
- Review
- Multiplex CRISPR-Cas Genome Editing: Next-Generation Microbial Strain Engineering.Journal of agricultural and food chemistry · 2024Review
- Expanding the CRISPR Toolbox for Engineering Lycopene Biosynthesis inMicroorganisms · 2024Article
- HyCas9-12aGEP: an efficient genome editing platform forFrontiers in bioengineering and biotechnology · 2024Article
- CRISPR-based gene editing technology and its application in microbial engineering.Engineering microbiology · 2023Review
- High-Yield Natural Vanillin Production byACS omega · 2023Article
- Novel configurations of type I-E CRISPR-Cas system in Corynebacterium striatum clinical isolates.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2023Article
- Development of a Transcriptional Factor PuuR-Based Putrescine-Specific Biosensor inBioengineering (Basel, Switzerland) · 2023Article
- 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
- CRISPR-assisted rational flux-tuning and arrayed CRISPRi screening of an L-proline exporter for L-proline hyperproduction.Nature communications · 2022Article
- Towards application of CRISPR-Cas12a in the design of modern viral DNA detection tools (Review).Journal of nanobiotechnology · 2022Review
- The application of CRISPR /Cas mediated gene editing in synthetic biology: Challenges and optimizations.Frontiers in bioengineering and biotechnology · 2022Review
- Identification of phage recombinase function unit in genus Corynebacterium.Applied microbiology and biotechnology · 2021Article
- Construction and application of a CRISPR/Cas9-assisted genomic editing system for Corynebacterium glutamicum.AMB Express · 2021Article
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8 authors.
Funding
Abstract
Corynebacterium glutamicum is an essential industrial strain that has been widely harnessed for the production of all kinds of value-added products. Efficient multiplex gene editing and large DNA fragment deletion are essential strategies for industrial biotechnological research. Cpf1 is a robust and simple genome editing tool for simultaneous editing of multiplex genes. However, no studies on effective multiplex gene editing and large DNA fragment deletion by the CRISPR/Cpf1 system in C. glutamicum have been reported. Here, we developed a multiplex gene editing method by optimizing the CRISPR/Cpf1-RecT system and a large chromosomal fragment deletion strategy using the CRISPR/Cpf1-RecET system in C. glutamicum ATCC 14067. The CRISPR/Cpf1-RecT system exhibited a precise editing efficiency of more than 91.6% with the PAM sequences TTTC, TTTG, GTTG or CTTC. The sites that could be edited were limited due to the PAM region and the 1-7 nt at the 5' end of the protospacer region. Mutations in the PAM region increased the editing efficiency of the - 6 nt region from 0 to 96.7%. Using a crRNA array, two and three genes could be simultaneously edited in one step via the CRISPR/Cpf1-RecT system, and the efficiency of simultaneously editing two genes was 91.6%, but the efficiency of simultaneously editing three genes was below 10%. The editing efficiency for a deletion of 1 kb was 79.6%, and the editing efficiencies for 5- and 20 kb length DNA fragment deletions reached 91.3% and 36.4%, respectively, via the CRISPR/Cpf1-RecET system. This research provides an efficient and simple tool for C. glutamicum genome editing that can further accelerate metabolic engineering efforts and genome evolution.
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