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Massively Parallel Profiling of RNA-targeting CRISPR-Cas13d
Hung-Che Kuo1, Joshua Prupes1, Chia-Wei Chou1
1Department of Molecular Biosciences and Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, Texas 78712, USA.
Biorxiv : the Preprint Server for Biology
|April 10, 2023
Summary
Researchers developed RNA-CHAMP to understand CRISPR-Cas13d RNA binding. This platform reveals Cas13d
Area of Science:
- Molecular Biology
- Biotechnology
- Genomics
Background:
- Type VI CRISPR enzymes are vital for gene regulation and diagnostics.
- A comprehensive understanding of their RNA binding specificity and cleavage activation is currently lacking.
Conclusions:
- A biophysical model indicates target recognition by Cas13d starts at the distal end of unstructured RNAs and proceeds proximally.
- Designed mismatched guide RNAs to modulate Cas13d activity for detecting single nucleotide polymorphisms (SNPs) in SARS-CoV-2 variants.
- RNA-CHAMP provides a quantitative method for studying protein-RNA interactions, enhancing CRISPR diagnostics and in vivo RNA editing.
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