Precise RNA targeting with CRISPR-Cas13d.

Hart, Sydney K; Müller, Simon; Wessels, Hans-Hermann; Méndez-Mancilla, Alejandro; Drabavicius, Gediminas; Choi, Olivia; Sanjana, Neville E · Nat Biotechnol · 2026

basic_science · Level V

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Abstract

The possibility of collateral RNA degradation poses a concern for transcriptome perturbations and therapeutic applications using CRISPR-Cas13. We show that collateral activity only occurs with high RfxCas13d expression. Using low-copy RfxCas13d in transcriptome-scale and combinatorial pooled screens, we achieve high on-target knockdown without extensive collateral activity. Furthermore, analysis of a high-fidelity Cas13 variant suggests that its reduced collateral activity may be due to overall diminished nuclease capability.

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