ArticleScientific reports2016
Multigene disruption in undomesticated Bacillus subtilis ATCC 6051a using the CRISPR/Cas9 system.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 53 papers.
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Who cites it
53 citing papers in PubMed.
- An inducer-independent, single-plasmid CRISPR-Cas9 system for genome editing in Bacillus species.Applied microbiology and biotechnology · 2026Article
- Secretory Production of Mature Protein-Glutaminase from Bacteroides helcogenes in Bacillus subtilis.Applied biochemistry and biotechnology · 2026Article
- Toward food-grade production of the Bacteroides helcogenes protein-glutamine glutaminase with an optimized Bacillus subtilis strain.Applied microbiology and biotechnology · 2026Article
- A three-plasmid-containing CRISPR-Cas9 platform to engineerApplied and environmental microbiology · 2025Article
- Engineering Bacillus subtilis for high-value bioproduction: recent advances and applications.Microbial cell factories · 2025Review
- Importance of the 5' untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007.AMB Express · 2025Article
- Synthetic communities as a model for determining interactions between a biofertilizer chassis organism and native microbial consortia.The ISME journal · 2025Article
- CRISPR-Cas9-based genome-editing technologies in engineering bacteria for the production of plant-derived terpenoids.Engineering microbiology · 2024Review
- Article
- A Programmable CRISPR/Cas9 Toolkit Improves Lycopene Production in Bacillus subtilis.Applied and environmental microbiology · 2023Article
- CRISPR-Based Gene Editing inPharmaceuticals (Basel, Switzerland) · 2023Review
- Improvement of Bacilysin Production inJournal of microbiology and biotechnology · 2023Article
- Development and application of CRISPR-based genetic tools in Bacillus species and Bacillus phages.Journal of applied microbiology · 2022Review
- Development and application of a rapid all-in-one plasmid CRISPR-Cas9 system for iterative genome editing in Bacillus subtilis.Microbial cell factories · 2022Article
- Development and application of a fast and efficient CRISPR-based genetic toolkit in Bacillus amyloliquefaciens LB1ba02.Microbial cell factories · 2022Article
- A facile and robust T7-promoter-based high-expression of heterologous proteins in Bacillus subtilis.Bioresources and bioprocessing · 2022Article
- Enhanced extracellular α-amylase production in Brevibacillus choshinensis by optimizing extracellular degradation and folding environment.Journal of industrial microbiology & biotechnology · 2022Article
- Overexpression ofFrontiers in microbiology · 2022Article
- Keratinases as Versatile Enzymatic Tools for Sustainable Development.Biomolecules · 2021Review
- Enzymatic deamination of the epigenetic nucleoside N6-methyladenosine regulates gene expression.Nucleic acids research · 2021Article
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Bacillus subtilis ATCC 6051a is an undomesticated strain used in the industrial production of enzymes. Because it is poorly transformable, genetic manipulation in this strain requires a highly efficient genome editing method. In this study, a Streptococcus pyogenes CRISPR/Cas9 system consisting of an all-in-one knockout plasmid containing a target-specific guide RNA, cas9, and a homologous repair template was established for highly efficient gene disruption in B. subtilis ATCC 6051a. With an efficiency of 33% to 53%, this system was used to disrupt the srfC, spoIIAC, nprE, aprE and amyE genes of B. subtilis ATCC 6051a, which hamper its use in industrial fermentation. Compared with B. subtilis ATCC 6051a, the final mutant, BS5 (ΔsrfC, ΔspoIIAC, ΔnprE, ΔaprE, ΔamyE), produces much less foam during fermentation, displays greater resistant to spore formation, and secretes 2.5-fold more β-cyclodextrin glycosyltransferase into the fermentation medium. Thus, the CRISPR/Cas9 system proved to be a powerful tool for targeted genome editing in an industrially relevant, poorly transformable strain.
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