ArticleJournal of industrial microbiology & biotechnology2020
Fine-tuning the regulation of Cas9 expression levels for efficient CRISPR-Cas9 mediated recombination in Streptomyces.
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 26 papers.
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Who cites it
26 citing papers in PubMed.
- An efficient endogenous type I-E CRISPR-Cas genome-editing platform for producing transglutaminase inEngineering microbiology · 2026Article
- Transferable genetic toolsets for nonmodel gut Clostridia enable in vivo reversible control of metabolite production.Nature biotechnology · 2026Article
- Multi-omics analysis of interspecies interactions in a soil Streptomyces community provides functional insights into siderophore ecology.Scientific reports · 2026Article
- Synthetic Biology Strategies for Activating Cryptic BGCs inACS omega · 2026Review
- Enhancing conjugation from E. coli to Streptomyces coelicolor by incorporating traJ into mobilizable plasmids.Applied microbiology and biotechnology · 2025Article
- A CRISPR-Cas9 system for knock-out and knock-in of high molecular weight DNA enables module-swapping of the pikromycin synthase in its native host.Microbial cell factories · 2025Article
- Application of CRISPR-Cas9 in microbial cell factories.Biotechnology letters · 2025Review
- CASCADE-Cas3 enables highly efficient genome engineering in Streptomyces species.Nucleic acids research · 2025Article
- Engineered CRISPR-Cas9 for Streptomyces sp. genome editing to improve specialized metabolite production.Nature communications · 2025Article
- CRISPR/Cas genome editing in plants: mechanisms, applications, and overcoming bottlenecks.Functional & integrative genomics · 2024Review
- Genetic transformation of the frog-killing chytrid fungusProceedings of the National Academy of Sciences of the United States of America · 2024Article
- Manipulation and epigenetic control of silent biosynthetic pathways in actinobacteria.World journal of microbiology & biotechnology · 2024Review
- Development of a genetic engineering toolbox for syngas-utilizing acetogen Clostridium sp. AWRP.Microbial cell factories · 2024Article
- Ablation ofPNAS nexus · 2023Article
- A generalizable Cas9/sgRNA prediction model using machine transfer learning with small high-quality datasets.Nature communications · 2023Article
- CRISPR/Cas9-mediated genome editing in vancomycin-producing strainFrontiers in bioengineering and biotechnology · 2023Article
- Characterization of the Biosynthetic Gene Cluster and Shunt Products Yields Insights into the Biosynthesis of Balmoralmycin.Applied and environmental microbiology · 2022Article
- Inducible CRISPR/Cas9 Allows for Multiplexed and Rapidly Segregated Single-Target Genome Editing inACS synthetic biology · 2022Article
- Two strategies to improve the supply of PKS extender units for ansamitocin P-3 biosynthesis by CRISPR-Cas9.Bioresources and bioprocessing · 2022Article
- CRISPR-Mediated Base Editing: From Precise Point Mutation to Genome-Wide Engineering in Nonmodel Microbes.Biology · 2022Review
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Authors and funding
4 authors.
Funding
Abstract
CRISPR-Cas9 has proven as a very powerful gene editing tool for Actinomyces, allowing scarless and precise genome editing in selected strains of these biotechnologically relevant microorganisms. However, its general application in actinomycetes has been limited due to its inefficacy when applying the system in an untested strain. Here, we provide evidence of how Cas9 levels are toxic for the model actinomycetes Streptomyces coelicolor M145 and Streptomyces lividans TK24, which show delayed or absence of growth. We overcame this toxicity by lowering Cas9 levels and have generated a set of plasmids in which Cas9 expression is either controlled by theophylline-inducible or constitutive promoters. We validated the targeting of these CRISPR-Cas9 system using the glycerol uptake operon and the actinorhodin biosynthesis gene cluster. Our results highlight the importance of adjusting Cas9 expression levels specifically in strains to gain optimum and efficient gene editing in Actinomyces.
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