DNA-guided CRISPR-Cas12 for cellular RNA targeting - PubMed
3 days ago
- #RNA targeting
- #CRISPR-Cas12
- #epitranscriptomic editing
- Development of a DNA-based guide (DNA) enabling Cas12 nucleases to target RNA, overcoming reliance on RNA-guided systems.
- Engineering of DNA to mimic a reverse-oriented CRISPR RNA scaffold, allowing AsCas12a and Cas12i1 to recognize RNA and trigger ssDNA trans-cleavage for sensitive RNA detection.
- Demonstrated 100% accuracy in hepatitis C virus RNA detection from clinical samples using DNA-guided Cas12 systems.
- Achieved 70-95% multiplex knockdown of endogenous RNA transcripts through ribosome stalling in multiple human cell lines.
- Mechanistic studies show activity depends on a stem loop stabilizing the Cas12-DNA-RNA complex.
- Codelivery of crRNA and DNA enables simultaneous DNA editing and RNA knockdown with a single effector.
- Modular fusions extend DNA applications to RNase H-mediated RNA degradation and METTL3-based epitranscriptomic editing.