Targeted RNA degradation remains a challenge using existing methods owing to off-target effects and toxicity. We adapted the CRISPR-Csm complex, a multi-protein effector from type III CRISPR systems, for precise knockdown of nuclear or cytoplasmic transcripts in eukaryotic cells with minimal side effects.
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References
Lennox, K. A. & Behlke, M. A. Mini-review on current strategies to knockdown long non-coding RNAs. J. Rare Dis. Res. Treat. 1, 66–70 (2016). A review article that describes the limitations of RNAi, especially for knockdown of long non-coding RNAs.
Ai, Y., Liang, D. & Wilusz, J. E. CRISPR/Cas13 effectors have differing extents of off-target effects that limit their utility in eukaryotic cells. Nucleic Acids Res. 50, e65 (2022). This paper reports the toxic side effects of Cas13’s trans-cleavage when used for RNA knockdown in eukaryotic cells.
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This is a summary of: Colognori, D., Trinidad, M. & Doudna, J. A. Precise transcript targeting by CRISPR-Csm complexes. Nat. Biotechnol. https://doi.org/10.1038/s41587-022-01649-9 (2023).
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CRISPR-Csm for eukaryotic RNA knockdown and imaging without toxicity. Nat Biotechnol 41, 1204–1205 (2023). https://doi.org/10.1038/s41587-023-01665-3
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DOI: https://doi.org/10.1038/s41587-023-01665-3
