Chemically modified guide RNAs enhance CRISPR-Cas13 knockdown in human cells.
Chemically modified guide RNAs enhance CRISPR-Cas13 knockdown in human cells.
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DOI：
10.1016/j.chembiol.2021.07.011
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发表时间：
2022-02-17
影响因子：
8.6
通讯作者：
Sanjana NE
中科院分区：
文献类型：
--
作者：
Méndez-Mancilla A;Wessels HH;Legut M;Kadina A;Mabuchi M;Walker J;Robb GB;Holden K;Sanjana NE
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. By screening multiple chemical modifications of the CRISPR RNA (crRNA), Méndez-Mancilla et al. identify specific chemical modifications and placement of modified bases that improve CRISPR-Cas13 transcript knockdown in human cells. The authors demonstrate that Cas13 ribonucleoproteins complexed with chemically modified crRNAs can modify the transcriptome of human primary T cells
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影响因子：
16.6
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通讯作者：
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11.8
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通讯作者：
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影响因子：
16.6
作者：
Barnes, Kayla G.;Lachenauer, Anna E.;Sabeti, Pardis C.
通讯作者：
Sabeti, Pardis C.