Exapted CRISPR-Cas12f homologues drive RNA-guided transcription - PubMed
11 hours ago
- #Transcription
- #Gene Activation
- #CRISPR
- Bacterial transcription initiation is tightly regulated by sigma (σ) factors.
- Bacteroidetes species encode dozens of specialized σE factors with unknown roles.
- A mechanism of RNA-guided gene activation involves σE factor and nuclease-dead Cas12f (dCas12f).
- dCas12f and σE homologues in E. coli show robust guide RNA enrichment and DNA target binding.
- Recruitment of σE depends on dCas12f and guide RNA, suggesting direct protein-protein interactions.
- The dCas12f-gRNA-σE ternary complex can recruit RNAP holoenzyme for gene expression.
- dCas12f-RNA-σE complexes drive gene expression without requisite promoter motifs.
- Transcription start sites are defined by the distance from the dCas12f-mediated R-loop.
- This system resembles CRISPR activation (CRISPRa) technology.