ArticleMicrobial biotechnology2021
A plasmid toolset for CRISPR-mediated genome editing and CRISPRi gene regulation in Escherichia coli.
Article in Microbial biotechnology, 2021. 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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Who cites it
17 citing papers in PubMed, 30 citations in OpenAlex.
- Conditional dCas12-Mediated Gene Knockdown and Complementation in Chlamydia.Methods in molecular biology (Clifton, N.J.) · 2027Article
- Prospects and challenges in using engineered lactic acid bacteria in aquaculture applications.Engineering microbiology · 2026Review
- Functional characterization of a type I-F1 CRISPR-cas system from the clinical isolate Shewanella xiamenensis Sh95 reveals constitutive activity and plasmid-curing capability.Scientific reports · 2026Article
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- A lineage-specific heat-induced feedback loop controls HrcA to promote chlamydial fitness under stress.bioRxiv : the preprint server for biology · 2025Article
- A rapid and efficient strategy for combinatorial repression of multiple genes in Escherichia coli.Microbial cell factories · 2025Article
- A parallel bioreactor strategy to rapidly determine growth-coupling relationships for bioproduction: a mevalonate case study.Biotechnology for biofuels and bioproducts · 2025Article
- Engineering an Escherichia coli strain for enhanced production of flavonoids derived from pinocembrin.Microbial cell factories · 2024Article
- A growth-coupling strategy for improving the stability of terpenoid bioproduction in Escherichia coli.Microbial cell factories · 2024Article
- EcCas6e-based antisense crRNA for gene repression and RNA editing in microorganisms.Nucleic acids research · 2024Article
- BacPE: a versatile prime-editing platform in bacteria by inhibiting DNA exonucleases.Nature communications · 2024Article
- Application of functional genomics for domestication of novel non-model microbes.Journal of industrial microbiology & biotechnology · 2024Review
- Carboxylic acid reductase-dependent biosynthesis of eugenol and related allylphenols.Microbial cell factories · 2023Article
- Research progress on nucleic acid detection and genome editing of CRISPR/Cas12 system.Molecular biology reports · 2023Review
- Bioproduction of Linalool From Paper Mill Waste.Frontiers in bioengineering and biotechnology · 2022Article
- A plasmid toolset for CRISPR-mediated genome editing and CRISPRi gene regulation in Escherichia coli.Microbial biotechnology · 2021Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
Abstract
CRISPR technologies have become standard laboratory tools for genetic manipulations across all kingdoms of life. Despite their origins in bacteria, the development of CRISPR tools for engineering bacteria has been slower than for eukaryotes; nevertheless, their function and application for genome engineering and gene regulation via CRISPR interference (CRISPRi) has been demonstrated in various bacteria, and adoption has become more widespread. Here, we provide simple plasmid-based systems for genome editing (gene knockouts/knock-ins, and genome integration of large DNA fragments) and CRISPRi in E. coli using a CRISPR-Cas12a system. The described genome engineering protocols allow markerless deletion or genome integration in just seven working days with high efficiency (> 80% and 50%, respectively), and the CRISPRi protocols allow robust transcriptional repression of target genes (> 90%) with a single cloning step. The presented minimized plasmids and their associated design and experimental protocols provide efficient and effective CRISPR-Cas12 genome editing, genome integration and CRISPRi implementation. These simple-to-use systems and protocols will allow the easy adoption of CRISPR technology by any laboratory.
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