ArticleNucleic acids research2022
A versatile Cas12k-based genetic engineering toolkit (C12KGET) for metabolic engineering in genetic manipulation-deprived strains.
Article in Nucleic acids research, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed, 31 citations in OpenAlex.
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- A Practical CRISPR-Based Method for Rapid Genome Editing inBio-protocol · 2025Article
- Advances in large-scale DNA engineering with the CRISPR system.Experimental & molecular medicine · 2025Review
- A CRISPR-SpCas9M-reporting system for efficient and rapid genome editing in Caulobacter crescentus.Nucleic acids research · 2025Article
- Concentration-Dependent CsrA Regulation of theACS synthetic biology · 2025Article
- Genomic engineering inFrontiers in microbiology · 2025Article
- Recent Advances in Genetic Engineering Strategies ofACS synthetic biology · 2024Review
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- Next-generation CRISPR technology for genome, epigenome and mitochondrial editing.Transgenic research · 2024Review
- A hybrid RNA-protein biosensor for high-throughput screening of adenosylcobalamin biosynthesis.Synthetic and systems biotechnology · 2024Article
- CRISPR/Cpf1-FOKI-induced gene editing inSynthetic and systems biotechnology · 2024Article
- Advances in miniature CRISPR-Cas proteins and their applications in gene editing.Archives of microbiology · 2024Review
- Recent application of CRISPR-Cas12 and OMEGA system for genome editing.Molecular therapy : the journal of the American Society of Gene Therapy · 2024Review
- Effect of blocking the haem synthesis pathway and weakening the haem synthesis pathway for sirohaem on the growth of and vitamin BBioprocess and biosystems engineering · 2023Article
- A type I-F CRISPRi system unveils the novel role of CzcR in modulating multidrug resistance ofMicrobiology spectrum · 2023Article
- A synthetic cell-free 36-enzyme reaction system for vitamin BNature communications · 2023Article
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The genetic modification of microorganisms is conducive to the selection of high-yield producers of high-value-added chemicals, but a lack of genetic tools hinders the industrialization of most wild species. Therefore, it is crucial to develop host-independent gene editing tools that can be used for genetic manipulation-deprived strains. The Tn7-like transposon from Scytonema hofmanni has been shown to mediate homologous recombination-independent genomic integration after heterologous expression in Escherichia coli, but the integration efficiency of heterologous sequences larger than 5 kb remains suboptimal. Here, we constructed a versatile Cas12k-based genetic engineering toolkit (C12KGET) that can achieve genomic integration of fragments up to 10 kb in size with up to 100% efficiency in challenging strains. Using C12KGET, we achieved the first example of highly efficient genome editing in Sinorhizobium meliloti, which successfully solved the problem that industrial strains are difficult to genetically modify, and increased vitamin B12 production by 25%. In addition, Cas12k can be directly used for transcriptional regulation of genes with up to 92% efficiency due to its naturally inactivated nuclease domain. The C12KGET established in this study is a versatile and efficient marker-free tool for gene integration as well as transcriptional regulation that can be used for challenging strains with underdeveloped genetic toolkits.
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