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化学修饰的引导RNA增强了人类原代细胞中的CRISPR-Cas基因组编辑。

Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells.

作者信息

Hendel Ayal, Bak Rasmus O, Clark Joseph T, Kennedy Andrew B, Ryan Daniel E, Roy Subhadeep, Steinfeld Israel, Lunstad Benjamin D, Kaiser Robert J, Wilkens Alec B, Bacchetta Rosa, Tsalenko Anya, Dellinger Douglas, Bruhn Laurakay, Porteus Matthew H

机构信息

Department of Pediatrics, Stanford University, Stanford, California, USA.

Agilent Research Laboratories, Santa Clara, California, USA.

出版信息

Nat Biotechnol. 2015 Sep;33(9):985-989. doi: 10.1038/nbt.3290. Epub 2015 Jun 29.

DOI:10.1038/nbt.3290
PMID:26121415
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4729442/
Abstract

CRISPR-Cas-mediated genome editing relies on guide RNAs that direct site-specific DNA cleavage facilitated by the Cas endonuclease. Here we report that chemical alterations to synthesized single guide RNAs (sgRNAs) enhance genome editing efficiency in human primary T cells and CD34(+) hematopoietic stem and progenitor cells. Co-delivering chemically modified sgRNAs with Cas9 mRNA or protein is an efficient RNA- or ribonucleoprotein (RNP)-based delivery method for the CRISPR-Cas system, without the toxicity associated with DNA delivery. This approach is a simple and effective way to streamline the development of genome editing with the potential to accelerate a wide array of biotechnological and therapeutic applications of the CRISPR-Cas technology.

摘要

CRISPR-Cas介导的基因组编辑依赖于引导RNA，其通过Cas核酸内切酶促进位点特异性DNA切割。我们在此报告，对合成的单向导RNA（sgRNA）进行化学修饰可提高人类原代T细胞和CD34(+)造血干细胞及祖细胞中的基因组编辑效率。将化学修饰的sgRNA与Cas9 mRNA或蛋白质共同递送，是一种基于RNA或核糖核蛋白（RNP）的高效CRISPR-Cas系统递送方法，且无DNA递送相关的毒性。这种方法是简化基因组编辑开发的简单有效方式，有可能加速CRISPR-Cas技术的广泛生物技术和治疗应用。

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