Defining and improving the genome-wide specificities of CRISPR-Cas9 nucleases.

Defining and improving the genome-wide specificities of CRISPR-Cas9 nucleases.
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DOI:
10.1038/nrg.2016.28
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发表时间:
2016-05
期刊:
Nature reviews. Genetics
影响因子:
--
通讯作者:
Joung JK
Joung JK
中科院分区:
其他
文献类型:
--
作者:
Tsai SQ;Joung JK

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CRISPR-Cas9 RNA引导的核酸酶是一种对生物学、遗传学和医学具有变革性的技术,因为它们可以简单地编程来切割活细胞和生物体中的特定DNA靶点。然而,为了将这些强大的分子工具转化为安全、有效的临床应用,仔细定义和改进它们在基因组范围内的特异性将是至关重要的。在这里,我们概述了我们对CRISPR-Cas9核酸酶识别和切割目标DNA的最新了解,确定和改进其特异性的方法,以及如何评估和减少研究和治疗应用中的非靶点x效应的关键考虑因素。
CRISPR-Cas9 RNA-guided nucleases are a transformative technology for biology, genetics, and medicine due to the simplicity with which they can be programmed to cleave specific DNA target sites in living cells and organisms. However, to translate these powerful molecular tools into safe, effective clinical applications, it will be of critical importance to carefully define and improve their genome-wide specificities. Here we outline our state-of-the-art understanding of target DNA recognition and cleavage by CRISPR-Cas9 nucleases, methods to determine and improve their specificities, and key considerations for how to evaluate and reduce off-target x effects for research and therapeutic applications.
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