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RNA-Responsive gRNAs for Controlling CRISPR Activity: Current Advances, Future Directions, and Potential Applications
Oana Pelea1, Tudor A Fulga1, Tatjana Sauka-Spengler1,2
1Radcliffe Department of Medicine, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom; and Kansas City, Missouri, USA.
The CRISPR Journal
|October 7, 2022
Summary
RNA-responsive CRISPR systems offer precise genome editing by sensing cellular RNAs. Engineering single-guide RNAs (sgRNAs) enhances control, paving the way for advanced therapeutic and research applications.
Area of Science:
- Molecular Biology
- Genome Engineering
- Biotechnology
Background:
- CRISPR-Cas9 is a powerful genome editing tool but can cause off-target effects.
- Controlling CRISPR expression is crucial for safety and specificity.
- Endogenous RNA levels reflect cellular states, offering a target for responsive systems.
Purpose of the Study:
- To review engineering strategies for RNA-responsive CRISPR systems.
- To focus on modifying *Streptococcus pyogenes* single-guide RNAs (sgRNAs).
- To discuss recent advances and future applications of RNA-responsive sgRNAs.
Main Methods:
- Engineering of single-guide RNA (sgRNA) structures.
- Development of systems responsive to endogenous RNA expression levels.
- Focus on modifications applicable to *Streptococcus pyogenes* CRISPR-Cas9 systems.
Main Results:
- sgRNA engineering is key to developing RNA-responsive CRISPR systems.
- RNA-sensing CRISPR offers precise control over genome editing.
- Optimized systems hold potential for novel applications.
Conclusions:
- RNA-responsive CRISPR technologies are advancing rapidly.
- Further optimization of sgRNAs can unlock new therapeutic and research avenues.
- This approach enables sophisticated control over gene editing based on cellular RNA signals.
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