Correction of a Genetic Disease in Mouse via Use of CRISPR-Cas9
Correction of a Genetic Disease in Mouse via Use of CRISPR-Cas9
复制标题
DOI：
10.1016/j.stem.2013.10.016
复制
发表时间：
2013-12-05
期刊：
影响因子：
23.9
通讯作者：
Li, Jinsong
中科院分区：
文献类型：
--
作者：
Wu, Yuxuan;Liang, Dan;Li, Jinsong
CRISPR-Cas9系统已被用于在一系列不同的生物体中产生突变等位基因。然而，到目前为止，还没有报道使用该系统来有效校正遗传疾病。在这里，我们表明，可以通过向受精卵中共注射Cas9 mRNA和针对突变等位基因的单向导RNA（sgRNA）来拯救Crygc基因显性突变导致白内障的小鼠。基于外源性提供的寡核苷酸或内源性WT等位基因，通过同源性定向修复（HDR）进行校正，仅有罕见的脱靶修饰证据。由此产生的小鼠是可生育的，并且能够将校正的等位基因传递给它们的后代。因此，我们的研究为使用CRISPR-Cas9系统纠正遗传性疾病提供了原理证明。
The CRISPR-Cas9 system has been employed to generate mutant alleles in a range of different organisms. However, so far there have not been reports of use of this system for efficient correction of a genetic disease. Here we show that mice with a dominant mutation in Crygc gene that causes cataracts could be rescued by coinjection into zygotes of Cas9 mRNA and a single-guide RNA (sgRNA) targeting the mutant allele. Correction occurred via homology-directed repair (HDR) based on an exogenously supplied oligonucleotide or the endogenous WT allele, with only rare evidence of off-target modifications. The resulting mice were fertile and able to transmit the corrected allele to their progeny. Thus, our study provides proof of principle for use of the CRISPR-Cas9 system to correct genetic disease.