ArticleNucleic acids research2022
CRISPRactivation-SMS, a message for PAM sequence independent gene up-regulation in Escherichia coli.
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 10 papers.
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Who cites it
10 citing papers in PubMed, 20 citations in OpenAlex.
- Metabolite-responsive scaffold RNAs for dynamic CRISPR transcriptional regulation.Nucleic acids research · 2025Article
- Site-Directed Genome Integration via Recombinase-Mediated Cassette Exchange (RMCE) inACS synthetic biology · 2025Article
- Genome-Wide CRISPRi Screening of Key Genes for Recombinant Protein Expression in Bacillus Subtilis.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Modifications of the 5' region of the CASPONMicrobial cell factories · 2024Article
- 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
- Cas9 off-target binding to the promoter of bacterial genes leads to silencing and toxicity.Nucleic acids research · 2023Article
- CRISPR/FnCas12a-mediated efficient multiplex and iterative genome editing in bacterial plant pathogens without donor DNA templates.PLoS pathogens · 2023Article
- Expanding the Scope of Bacterial CRISPR Activation with PAM-Flexible dCas9 Variants.ACS synthetic biology · 2022Article
- From DNA-protein interactions to the genetic circuit design using CRISPR-dCas systems.Frontiers in molecular biosciences · 2022Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Governance of the endogenous gene regulatory network enables the navigation of cells towards beneficial traits for recombinant protein production. CRISPRactivation and interference provides the basis for gene expression modulation but is primarily applied in eukaryotes. Particularly the lack of wide-ranging prokaryotic CRISPRa studies might be attributed to intrinsic limitations of bacterial activators and Cas9 proteins. While bacterial activators need accurate spatial orientation and distancing towards the target promoter to be functional, Cas9-based CRISPR tools only bind sites adjacent to NGG PAM sequences. These circumstances hampered Cas9-guided activators from mediating the up-regulation of endogenous genes at precise positions in bacteria. We could overcome this limitation by combining the PAM independent Cas9 variant SpRY and a CRISPRa construct using phage protein MCP fused to transcriptional activator SoxS. This CRISPRa construct, referred to as SMS, was compared with previously reported CRISPRa constructs and showed up-regulation of a reporter gene library independent of its PAM sequence in Escherichia coli. We also demonstrated down-regulation and multi-gene expression control with SMS at non-NGG PAM sites. Furthermore, we successfully applied SMS to up-regulate endogenous genes, and transgenes at non-NGG PAM sites, which was impossible with the previous CRISPRa construct.
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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.