ArticleNature protocols2020
CRISPR-Cas9, CRISPRi and CRISPR-BEST-mediated genetic manipulation in streptomycetes.
Article in Nature protocols, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers.
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Who cites it
46 citing papers in PubMed, 92 citations in OpenAlex.
- Expanding perspectives on specialized metabolism in Streptomyces bacteria.Essays in biochemistry · 2026Review
- Inducible CRISPRi enables efficient and high-fidelity genome editing in Streptomyces.Nucleic acids research · 2026Article
- Molecular landscape analysis of azinothricin-type natural products enables the identification of kettapeptin fromSynthetic and systems biotechnology · 2026Article
- Multi-omics analysis of interspecies interactions in a soil Streptomyces community provides functional insights into siderophore ecology.Scientific reports · 2026Article
- CRISPy-web 3.0: A unified platform for multi-modal guide RNA design for CRISPR and TnpB genome editing applications.Synthetic and systems biotechnology · 2026Article
- Technological advances in axonal regeneration and structural plasticity: optogenetics, single-cell omics, and tissue clearing.Frontiers in cellular neuroscience · 2026Review
- StreptoCAD: An Open-Source Software Toolbox Automating Genome Engineering Workflows in Streptomycetes.ACS synthetic biology · 2025Article
- Breaking down microplastics: insights into the role of actinomycetes in biotic degradation pathways.Archives of microbiology · 2025Review
- P450 Enzyme LyoI Performs Hydro-2,2'-Bifuran Oxidation in the Polyether Ionophore Lysocellin.Angewandte Chemie (International ed. in English) · 2025Article
- STAGE: A compact and versatile TnpB-based genome editing toolkit forProceedings of the National Academy of Sciences of the United States of America · 2025Article
- ActinoMation: A literate programming approach for medium-throughput robotic conjugation ofSynthetic and systems biotechnology · 2025Article
- Identification of a critical gene involved in the biosynthesis of the polyene macrolide lavencidin inApplied and environmental microbiology · 2025Article
- Structural Characterisation of TetR/AcrR Regulators inInternational journal of molecular sciences · 2025Article
- Evolution-Guided Discovery of Antimycobacterial Triculamin-Like Lasso Peptides.Angewandte Chemie (International ed. in English) · 2025Article
- Genome Mining Reveals Rifamycin Biosynthesis in a Taklamakan Desert Actinomycete.Microorganisms · 2025Article
- Make-or-break prime editing for genome engineering in Streptococcus pneumoniae.Nature communications · 2025Article
- The Biarylitides: Understanding the Structure and Biosynthesis of a Fascinating Class of Cytochrome P450 Modified RiPP Natural Products.Chembiochem : a European journal of chemical biology · 2025Review
- CASCADE-Cas3 enables highly efficient genome engineering in Streptomyces species.Nucleic acids research · 2025Article
- A rational multi-target combination strategy for synergistic improvement of non-ribosomal peptide production.Nature communications · 2025Article
- Engineered CRISPR-Cas9 for Streptomyces sp. genome editing to improve specialized metabolite production.Nature communications · 2025Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Streptomycetes are prominent sources of bioactive natural products, but metabolic engineering of the natural products of these organisms is greatly hindered by relatively inefficient genetic manipulation approaches. New advances in genome editing techniques, particularly CRISPR-based tools, have revolutionized genetic manipulation of many organisms, including actinomycetes. We have developed a comprehensive CRISPR toolkit that includes several variations of 'classic' CRISPR-Cas9 systems, along with CRISPRi and CRISPR-base editing systems (CRISPR-BEST) for streptomycetes. Here, we provide step-by-step protocols for designing and constructing the CRISPR plasmids, transferring these plasmids to the target streptomycetes, and identifying correctly edited clones. Our CRISPR toolkit can be used to generate random-sized deletion libraries, introduce small indels, generate in-frame deletions of specific target genes, reversibly suppress gene transcription, and substitute single base pairs in streptomycete genomes. Furthermore, the toolkit includes a Csy4-based multiplexing option to introduce multiple edits in a single experiment. The toolkit can be easily extended to other actinomycetes. With our protocol, it takes <10 d to inactivate a target gene, which is much faster than alternative protocols.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.