CRISPR/Cas9 mediated genome engineering in Drosophila
CRISPR/Cas9 mediated genome engineering in Drosophila
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DOI：
10.1016/j.ymeth.2014.02.019
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发表时间：
2014-09-01
期刊：
影响因子：
4.8
通讯作者：
Liu, Ji-Long
中科院分区：
文献类型：
--
作者：
Bassett, Andrew;Liu, Ji-Long
基因组工程已经彻底改变了许多生物的基因分析。在这里，我们描述了一种简单而有效的技术来产生和检测在黑腹果蝇所需的靶基因的新突变。我们通过向果蝇胚胎注射编码Cas9内切酶的mRNA和体外转录的合成引导RNA，将双链断裂定位到基因组内的特定位点。通过对整个果蝇和单个翅膀的高分辨率熔体分析，可以检测到由于该位点非同源末端连接低效而导致的小插入和删除突变，从而在1个月内获得稳定的系。(C) 2014年由Elsevier Inc.出版。
Genome engineering has revolutionised genetic analysis in many organisms. Here we describe a simple and efficient technique to generate and detect novel mutations in desired target genes in Drosophila melanogaster. We target double strand breaks to specific sites within the genome by injecting mRNA encoding the Cas9 endonuclease and in vitro transcribed synthetic guide RNA into Drosophila embryos. The small insertion and deletion mutations that result from inefficient non-homologous end joining at this site are detected by high resolution melt analysis of whole flies and individual wings, allowing stable lines to be made within 1 month. (C) 2014 Published by Elsevier Inc.