Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system.

Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system.
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DOI:
10.1038/srep05400
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发表时间:
2014-06-23
期刊:
影响因子:
4.6
通讯作者:
Yamamoto T
Yamamoto T
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Sakuma T;Nishikawa A;Kume S;Chayama K;Yamamoto T

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CRISPR/Cas9介导的基因组编辑是下一代遗传修饰策略,不仅用于单基因靶向,还用于多靶向诱变。为了充分利用CRISPR/Cas9的多重性,我们建立了一种用于构建包含多个指导RNA表达盒和Cas9核酸酶/切口酶表达盒的一体化表达载体的系统。我们进一步展示了使用我们的新系统构建的一体化CRISPR/Cas9载体在人类细胞中多次靶向(包括染色体缺失)的成功实例。我们的系统为多重基因组/表观基因组编辑、多个基因的同时激活/抑制等提供了有效的靶向策略。
CRISPR/Cas9-mediated genome editing is a next-generation strategy for genetic modifications, not only for single gene targeting, but also for multiple targeted mutagenesis. To make the most of the multiplexity of CRISPR/Cas9, we established a system for constructing all-in-one expression vectors containing multiple guide RNA expression cassettes and a Cas9 nuclease/nickase expression cassette. We further demonstrated successful examples of multiple targeting including chromosomal deletions in human cells using the all-in-one CRISPR/Cas9 vectors constructed with our novel system. Our system provides an efficient targeting strategy for multiplex genome/epigenome editing, simultaneous activation/repression of multiple genes, and beyond.
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