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利用CRISPR/Cas系统的多功能基因组工具

2013-01-20 10:04:45

浏览次数:4600

利用CRISPR/Cas系统的多功能基因组工具

来源:仪方生物 www.yeslab.com


遗传变异和元件的功能阐述需要精确的基因组编辑技术。2型原核CRISPR(规律成簇的间隔短回文重复)适应性免疫系统具有协助RNA介导的位点特异性DNA剪切。研究者设计了2个2型CRISPR系统并证实在人和小鼠细胞内源基因组位点Cas9核酸酶能够直接被短RNAs诱导进行精确剪切。Cas9也能够在最小突变活性的情况下被转化拟合酶来协助同源性定向修复。多能介导序列能够编码成单个CRISPR矩阵来刺激哺乳动物基因组的一些位点编辑,证明了该CRISPR技术的易设计和广泛的用途。

Multiplex Genome Engineering Using CRISPR/Cas Systems

Le Cong, F. Ann Ran, David Cox, Shuailiang Lin, Robert Barretto, Naomi Habib, Patrick D. Hsu, Xuebing Wu, Wenyan Jiang, Luciano A. Marraffini, Feng Zhang1

Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats) adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. We engineered two different type II CRISPR systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair with minimal mutagenic activity. Finally, multiple guide sequences can be encoded into a single CRISPR array to enable simultaneous editing of several sites within the mammalian genome, demonstrating easy programmability and wide applicability of the CRISPR technology.
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