ArticleNucleic acids research2021
Efficient multiplexed gene regulation in Saccharomyces cerevisiae using dCas12a.
Article in Nucleic acids research, 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, 39 citations in OpenAlex.
- CRISPR-based Transcriptional Regulation: Technologies, Applications, and Future Directions.DNA · 2025Article
- HyperdCas12a-based multiplexed genetic regulation in Candida albicans.Nucleic acids research · 2025Article
- Epigenome Engineering Using dCas Systems for Biomedical Applications and Biotechnology: Current Achievements, Opportunities and Challenges.International journal of molecular sciences · 2025Review
- Discovering the hidden function in fungal genomes.Nature communications · 2024Review
- The rise and future of CRISPR-based approaches for high-throughput genomics.FEMS microbiology reviews · 2024Review
- EcCas6e-based antisense crRNA for gene repression and RNA editing in microorganisms.Nucleic acids research · 2024Article
- Machine Learning-Guided Optimization ofACS synthetic biology · 2024Article
- Genome-scale CRISPRi screening: A powerful tool in engineering microbiology.Engineering microbiology · 2023Review
- CREEPY: CRISPR-mediated editing of synthetic episomes in yeast.Nucleic acids research · 2023Article
- Article
- Development and applications of a CRISPR activation system for facile genetic overexpression in Candida albicans.G3 (Bethesda, Md.) · 2023Article
- Shortened CRISPR-Cas9 arrays enable multiplexed gene targeting in bacteria from a smaller DNA footprint.RNA biology · 2023Article
- Recent Advances in Directed Yeast Genome Evolution.Journal of fungi (Basel, Switzerland) · 2022Review
- CRISPR-mediated protein-tagging signal amplification systems for efficient transcriptional activation and repression in Saccharomyces cerevisiae.Nucleic acids research · 2022Article
- Tips, Tricks, and Potential Pitfalls of CRISPR Genome Editing inFrontiers in bioengineering and biotechnology · 2022Review
- Comprehensive Genome Engineering Toolbox for MicroalgaeACS synthetic biology · 2021Article
- CRISPR-Based Genetic Switches and Other Complex Circuits: Research and Application.Life (Basel, Switzerland) · 2021Review
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 3 countries.
Funding
Abstract
CRISPR Cas12a is an RNA-programmable endonuclease particularly suitable for gene regulation. This is due to its preference for T-rich PAMs that allows it to more easily target AT-rich promoter sequences, and built-in RNase activity which can process a single CRISPR RNA array encoding multiple spacers into individual guide RNAs (gRNAs), thereby simplifying multiplexed gene regulation. Here, we develop a flexible dCas12a-based CRISPRi system for Saccharomyces cerevisiae and systematically evaluate its design features. This includes the role of the NLS position, use of repression domains, and the position of the gRNA target. Our optimal system is comprised of dCas12a E925A with a single C-terminal NLS and a Mxi1 or a MIG1 repression domain, which enables up to 97% downregulation of a reporter gene. We also extend this system to allow for inducible regulation via an RNAP II-controlled promoter, demonstrate position-dependent effects in crRNA arrays, and use multiplexed regulation to stringently control a heterologous β-carotene pathway. Together these findings offer valuable insights into the design constraints of dCas12a-based CRISPRi and enable new avenues for flexible and efficient gene regulation in S. cerevisiae.
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Registered trials
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