清脆的
Cas9
秀丽隐杆线虫
同源重组
生物
基因组编辑
基因组
遗传学
计算生物学
基因组工程
基因
DNA
作者
Daniel J. Dickinson,Jordan D. Ward,David J. Reiner,Bob Goldstein
出处
期刊:Nature Methods
[Springer Nature]
日期:2013-09-01
卷期号:10 (10): 1028-1034
被引量:862
摘要
CRISPR-Cas9–mediated cleavage is used to stimulate homologous recombination at specific target sites in the C. elegans genome, permitting flexible tagging and sequence modification of endogenous worm genes. Study of the nematode Caenorhabditis elegans has provided important insights in a wide range of fields in biology. The ability to precisely modify genomes is critical to fully realize the utility of model organisms. Here we report a method to edit the C. elegans genome using the clustered, regularly interspersed, short palindromic repeats (CRISPR) RNA-guided Cas9 nuclease and homologous recombination. We demonstrate that Cas9 is able to induce DNA double-strand breaks with specificity for targeted sites and that these breaks can be repaired efficiently by homologous recombination. By supplying engineered homologous repair templates, we generated gfp knock-ins and targeted mutations. Together our results outline a flexible methodology to produce essentially any desired modification in the C. elegans genome quickly and at low cost. This technology is an important addition to the array of genetic techniques already available in this experimentally tractable model organism.
科研通智能强力驱动
Strongly Powered by AbleSci AI