Fusion guide RNAs for orthogonal gene manipulation with Cas9 and Cpf1.

Fusion guide RNAs for orthogonal gene manipulation with Cas9 and Cpf1.
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DOI:
10.1038/s41467-017-01650-w
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发表时间:
2017-11-23
影响因子:
16.6
通讯作者:
Kim Y
Kim Y
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Kweon J;Jang AH;Kim DE;Yang JW;Yoon M;Rim Shin H;Kim JS;Kim Y

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细菌衍生的成簇规则间隔短回文重复序列(CRISPR)-Cas系统是基因组工程的有力工具。最近,除了Cas蛋白工程,还进行了指导RNA的改进,有助于拓宽CRISPR-Cas9系统的研究领域。在这里,我们开发了一种融合引导RNA(fgRNA),它与Cas9和Cpf 1蛋白一起发挥作用,以诱导人类细胞中的突变。此外,我们证明了fgRNA可以用于两种类型的Cas蛋白的多重基因组编辑和正交基因组操作。我们的研究结果表明,fgRNA可以用作进行多种基因操作的工具。Cas9和Cpf 1都适用于基因组工程,编辑和基因表达调控。在这里,作者设计了一种融合向导RNA,可以与两种蛋白质相互作用,进行多重和正交基因组操作。
The bacteria-derived clustered regularly interspaced short palindromic repeat (CRISPR)–Cas systems are powerful tools for genome engineering. Recently, in addition to Cas protein engineering, the improvement of guide RNAs are also performed, contributing to broadening the research area of CRISPR–Cas9 systems. Here we develop a fusion guide RNA (fgRNA) that functions with both Cas9 and Cpf1 proteins to induce mutations in human cells. Furthermore, we demonstrate that fgRNAs can be used in multiplex genome editing and orthogonal genome manipulation with two types of Cas proteins. Our results show that fgRNAs can be used as a tool for performing multiple gene manipulations. Cas9 and Cpf1 have both been adapted for genome engineering, editing and gene expression regulation. Here the authors design a fusion guide RNA that can interact with both proteins for multiple and orthogonal genome manipulation.
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