Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells.
Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells.
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DOI：
10.1038/nbt.3290
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发表时间：
2015-09
影响因子：
46.9
通讯作者：
Porteus MH
中科院分区：
文献类型：
--
作者：
Hendel A;Bak RO;Clark JT;Kennedy AB;Ryan DE;Roy S;Steinfeld I;Lunstad BD;Kaiser RJ;Wilkens AB;Bacchetta R;Tsalenko A;Dellinger D;Bruhn L;Porteus MH
CRISPR-Cas介导的基因组编辑依赖于指导由Cas内切核酸酶促进的位点特异性DNA切割的指导RNA。在这里，我们报告了对合成的单向导RNA（sgRNA）的化学改变增强了人类原代T细胞和CD 34+造血干细胞和祖细胞中的基因组编辑效率。共递送化学修饰的sgRNA与Cas9 mRNA或蛋白质是用于CRISPR-Cas系统的有效的基于RNA或核糖核蛋白（RNP）的递送方法，而没有与DNA递送相关的毒性。这种方法是简化基因组编辑开发的一种简单有效的方法，有可能加速CRISPR-Cas技术的广泛生物技术和治疗应用。
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.