Author: Méndez-Mancilla, Alejandro; Wessels, Hans-Hermann; Legut, Mateusz; Kadina, Anastasia; Mabuchi, Megumu; Walker, John; Robb, G. Brett; Holden, Kevin; Sanjana, Neville E.
Title: Chemically modified guide RNAs enhance CRISPR-Cas13 knockdown in human cells Cord-id: cegw80t1 Document date: 2021_5_14
ID: cegw80t1
Snippet: RNA-targeting CRISPR-Cas13 proteins have recently emerged as a powerful platform to transiently modulate gene expression outcomes. However, protein and CRISPR RNA (crRNA) delivery in human cells can be challenging and knockdown can be transient due to rapid crRNA degradation. Here we compare several chemical RNA modifications at different positions to identify synthetic crRNAs that improve RNA targeting efficiency and half-life in human cells. We show that co-delivery of modified crRNAs and reco
Document: RNA-targeting CRISPR-Cas13 proteins have recently emerged as a powerful platform to transiently modulate gene expression outcomes. However, protein and CRISPR RNA (crRNA) delivery in human cells can be challenging and knockdown can be transient due to rapid crRNA degradation. Here we compare several chemical RNA modifications at different positions to identify synthetic crRNAs that improve RNA targeting efficiency and half-life in human cells. We show that co-delivery of modified crRNAs and recombinant Cas13 enzyme in ribonucleoprotein (RNP) complexes enables transient gene expression modulation in primary CD4+ and CD8+ T-cells. This system represents a robust and efficient method to transiently modulate transcripts without genetic manipulation.
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