用于 Cas9 和 Cpf1 正交基因操作的融合向导 RNA
Fusion guide RNAs for orthogonal gene manipulation with Cas9 and Cpf1.
机构信息
Department of Biomedical Sciences, University of Ulsan College of Medicine, Asan Medical Center, Seoul, 05505, Republic of Korea.
Department of Chemistry, Seoul National University, Seoul, 08826, Republic of Korea.
出版信息
Nat Commun. 2017 Nov 23;8(1):1723. doi: 10.1038/s41467-017-01650-w.
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.
细菌衍生的成簇规律间隔短回文重复（CRISPR）-Cas 系统是基因组工程的强大工具。最近，除了 Cas 蛋白工程外，还对向导 RNA 进行了改进，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向导 RNA（fgRNA），它可以与 Cas9 和 Cpf1 蛋白一起在人类细胞中诱导突变。此外，我们证明 fgRNA 可用于两种 Cas 蛋白的多重基因组编辑和正交基因组操作。我们的结果表明，fgRNA 可作为执行多个基因操作的工具。
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