ArticleMicrobial cell factories2017
Development of a CRISPR/Cas9 genome editing toolbox for Corynebacterium glutamicum.
Article in Microbial cell factories, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 54 papers.
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Who cites it
54 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
- A thermostable Cas9-based genome editing system for thermophilic acetogenic bacteriumApplied and environmental microbiology · 2025Article
- Engineering of L-threonine and L-proline biosensors by directed evolution of transcriptional regulator SerR and application for high-throughput screening.Bioresources and bioprocessing · 2025Article
- Cas9-independent tracrRNA cytotoxicity inmicroLife · 2025Article
- CRISPR-Cas9-based genome-editing technologies in engineering bacteria for the production of plant-derived terpenoids.Engineering microbiology · 2024Review
- Recent Advances in Metabolic Engineering for the Biosynthesis of Phosphoenol Pyruvate-Oxaloacetate-Pyruvate-Derived Amino Acids.Molecules (Basel, Switzerland) · 2024Review
- 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
- Promising non-model microbial cell factories obtained by genome reduction.Frontiers in bioengineering and biotechnology · 2024Review
- HyCas9-12aGEP: an efficient genome editing platform forFrontiers in bioengineering and biotechnology · 2024Article
- Establishment of CRISPR-Cpf1-assisted gene editing tool and engineering of 4-hydroxyisoleucine biosynthesis in Corynebacterium glutamicum.World journal of microbiology & biotechnology · 2023Article
- CRISPR-Based Gene Editing inPharmaceuticals (Basel, Switzerland) · 2023Review
- Sustainable and high-level microbial production of plant hemoglobin in Corynebacterium glutamicum.Biotechnology for biofuels and bioproducts · 2023Article
- In Vivo Genome Editing in Type I and II Methanotrophs Using a CRISPR/Cas9 System.ACS synthetic biology · 2023Article
- Effects of down-regulation of ackA expression by CRISPR-dCpf1 on succinic acid production in Actinobacillus succinogenes.AMB Express · 2023Article
- Enhanced production of D-pantothenic acid in Corynebacterium glutamicum using an efficient CRISPR-Cpf1 genome editing method.Microbial cell factories · 2023Article
- Engineering allosteric inhibition of homoserine dehydrogenase by semi-rational saturation mutagenesis screening.Frontiers in bioengineering and biotechnology · 2023Article
- Synthetic biology tools for engineeringComputational and structural biotechnology journal · 2023Review
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6 authors.
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Abstract
backgroundCorynebacterium glutamicum is an important industrial workhorse and advanced genetic engineering tools are urgently demanded. Recently, the clustered regularly interspaced short palindromic repeats (CRISPR) and their CRISPR-associated proteins (Cas) have revolutionized the field of genome engineering. The CRISPR/Cas9 system that utilizes NGG as protospacer adjacent motif (PAM) and has good targeting specificity can be developed into a powerful tool for efficient and precise genome editing of C. glutamicum.
resultsHerein, we developed a versatile CRISPR/Cas9 genome editing toolbox for C. glutamicum. Cas9 and gRNA expression cassettes were reconstituted to combat Cas9 toxicity and facilitate effective termination of gRNA transcription. Co-transformation of Cas9 and gRNA expression plasmids was exploited to overcome high-frequency mutation of cas9, allowing not only highly efficient gene deletion and insertion with plasmid-borne editing templates (efficiencies up to 60.0 and 62.5%, respectively) but also simple and time-saving operation. Furthermore, CRISPR/Cas9-mediated ssDNA recombineering was developed to precisely introduce small modifications and single-nucleotide changes into the genome of C. glutamicum with efficiencies over 80.0%. Notably, double-locus editing was also achieved in C. glutamicum. This toolbox works well in several C. glutamicum strains including the widely-used strains ATCC 13032 and ATCC 13869.
conclusionsIn this study, we developed a CRISPR/Cas9 toolbox that could facilitate markerless gene deletion, gene insertion, precise base editing, and double-locus editing in C. glutamicum. The CRISPR/Cas9 toolbox holds promise for accelerating the engineering of C. glutamicum and advancing its application in the production of biochemicals and biofuels.
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