ArticleNucleic acids research2022
CRISPR-mediated protein-tagging signal amplification systems for efficient transcriptional activation and repression in Saccharomyces cerevisiae.
Article in Nucleic acids research, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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12 citing papers in PubMed, 26 citations in OpenAlex.
- CRISPR-DNA Polymerase Assisted Targeted Mutagenesis for Regulable Laboratory Evolution.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Matrix regulation: a plug-and-tune method for combinatorial regulation in Saccharomyces cerevisiae.Nature communications · 2025Article
- Programmable genome engineering and gene modifications for plant biodesign.Plant communications · 2025Review
- Epigenome Engineering Using dCas Systems for Biomedical Applications and Biotechnology: Current Achievements, Opportunities and Challenges.International journal of molecular sciences · 2025Review
- Unravelling fungal genome editing revolution: pathological and biotechnological application aspects.Archives of microbiology · 2025Review
- The advances in creating Crabtree-negative Saccharomyces cerevisiae and the application for chemicals biosynthesis.FEMS yeast research · 2025Review
- Development of a bacterial gene transcription activating strategy based on transcriptional activator positive feedback.Journal of advanced research · 2024Article
- An integrative database and its application for plant synthetic biology research.Plant communications · 2024Article
- Genome-scale transcriptional activation by non-homologous end joining-mediated integration in Yarrowia lipolytica.Biotechnology for biofuels and bioproducts · 2024Article
- Advances in the dynamic control of metabolic pathways inEngineering microbiology · 2023Review
- Overcoming the Limitations of CRISPR-Cas9 Systems inMicroorganisms · 2023Review
- An outlook to sophisticated technologies and novel developments for metabolic regulation in theFrontiers in bioengineering and biotechnology · 2023Review
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Saccharomyces cerevisiae is an important model eukaryotic microorganism and widely applied in fundamental research and the production of various chemicals. Its ability to efficiently and precisely control the expression of multiple genes is valuable for metabolic engineering. The clustered regularly interspaced short palindromic repeats (CRISPR)-mediated regulation enables complex gene expression programming; however, the regulation efficiency is often limited by the efficiency of pertinent regulators. Here, we developed CRISPR-mediated protein-tagging signal amplification system for simultaneous multiplexed gene activation and repression in S. cerevisiae. By introducing protein scaffolds (SPY and SunTag systems) to recruit multiple copies of regulators to different nuclease-deficient CRISPR proteins and design optimization, our system amplified gene regulation efficiency significantly. The gene activation and repression efficiencies reached as high as 34.9-fold and 95%, respectively, being 3.8- and 8.6-fold higher than those observed on the direct fusion of regulators with nuclease-deficient CRISPR proteins, respectively. We then applied the orthogonal bifunctional CRISPR-mediated transcriptional regulation system to regulate the expression of genes associated with 3-hydroxypropanoic acid production to deduce that CRISPR-associated regulator recruiting systems represent a robust method for simultaneously regulating multiple genes and rewiring metabolic pathways.
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