ArticleNucleic acids research2021
Rational engineering of a modular bacterial CRISPR-Cas activation platform with expanded target range.
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 19 papers.
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Who cites it
19 citing papers in PubMed, 42 citations in OpenAlex.
- Metabolite-responsive scaffold RNAs for dynamic CRISPR transcriptional regulation.Nucleic acids research · 2025Article
- Computational design of sequence-specific DNA-binding proteins.Nature structural & molecular biology · 2025Article
- Prime Editing Modification with FEN1 Improves F508del Variant Editing in theInternational journal of molecular sciences · 2025Article
- How many plasmids can bacteria carry? A synthetic biology perspective.Open biology · 2025Review
- Development of a CRISPR activation system for targeted gene upregulation in Synechocystis sp. PCC 6803.Communications biology · 2025Article
- CRISPR/Cas13X-assisted programmable and multiplexed translation regulation for controlled biosynthesis.Nucleic acids research · 2025Article
- Genome-Wide CRISPRi Screening of Key Genes for Recombinant Protein Expression in Bacillus Subtilis.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Integrating bacterial molecular genetics with chemical biology for renewed antibacterial drug discovery.The Biochemical journal · 2024Review
- The expanded CRISPR toolbox for constructing microbial cell factories.Trends in biotechnology · 2024Review
- Synthetic Gene Circuits Combining CRISPR Interference and CRISPR Activation inACS synthetic biology · 2023Article
- Engineering activatable promoters for scalable and multi-input CRISPRa/i circuits.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Group B Streptococcus Cas9 variants provide insight into programmable gene repression and CRISPR-Cas transcriptional effects.Communications biology · 2023Article
- Expanding the Scope of Bacterial CRISPR Activation with PAM-Flexible dCas9 Variants.ACS synthetic biology · 2022Article
- CRISPRactivation-SMS, a message for PAM sequence independent gene up-regulation in Escherichia coli.Nucleic acids research · 2022Article
- Activating natural product synthesis using CRISPR interference and activation systems in Streptomyces.Nucleic acids research · 2022Article
- From DNA-protein interactions to the genetic circuit design using CRISPR-dCas systems.Frontiers in molecular biosciences · 2022Review
- Reprogramming Microbial COFrontiers in bioengineering and biotechnology · 2022Review
- CRISPR-Based Approaches for Gene Regulation in Non-Model Bacteria.Frontiers in genome editing · 2022Review
- Competitive dCas9 binding as a mechanism for transcriptional control.Molecular systems biology · 2021Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR-Cas activator (CRISPRa) systems that selectively turn on transcription of a target gene are a potentially transformative technology for programming cellular function. While in eukaryotes versatile CRISPRa systems exist, in bacteria these systems suffer from a limited ability to activate different genes due to strict distance-dependent requirements of functional target binding sites, and require greater customization to optimize performance in different genetic and cellular contexts. To address this, we apply a rational protein engineering approach to create a new CRISPRa platform that is highly modular to allow for easy customization and has increased targeting flexibility through harnessing engineered Cas proteins. We first demonstrate that transcription activation domains can be recruited by CRISPR-Cas through noncovalent protein-protein interactions, which allows each component to be encoded on separate and easily interchangeable plasmid elements. We then exploit this modularity to rapidly screen a library of different activation domains, creating new systems with distinct regulatory properties. Furthermore, we demonstrate that by harnessing a library of circularly permuted Cas proteins, we can create CRISPRa systems that have different target binding site requirements, which together, allow for expanded target range.
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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.