CRISPR/Cas9 mediates efficient conditional mutagenesis in Drosophila.

CRISPR/Cas9 mediates efficient conditional mutagenesis in Drosophila.
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CRISPR/Cas9 在果蝇中介导有效的条件诱变。

DOI:
10.1534/g3.114.014159
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发表时间:
2014-09-05
期刊:
G3 (Bethesda, Md.)
影响因子:
--
通讯作者:
Gao G
Gao G
中科院分区:
其他
文献类型:
--
作者:
Xue Z;Wu M;Wen K;Ren M;Long L;Zhang X;Gao G

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现有的转基因RNA干扰(RNAi)方法通过对果蝇中靶mRNA进行可控沉默,极大地促进了功能基因组研究。尽管RNAi方法极其强大,但其效率低下的问题仍然存在。在此,我们开发了一种CRISPR/Cas9介导的条件性诱变系统,该系统将由Gal4/上游激活序列系统驱动的Cas9的组织特异性表达与多种普遍表达的向导RNA转基因相结合,以便在时空可控的方式下有效地使基因表达失活。此外,通过在转基因载体中包含针对单个基因的多个向导RNA,我们在特定组织中实现了高度的基因诱变。CRISPR/Cas9介导的条件性诱变系统为基因功能分析提供了一种简单有效的工具,并对现有的RNAi方法进行了补充。
Existing transgenic RNA interference (RNAi) methods greatly facilitate functional genome studies via controlled silencing of targeted mRNA in Drosophila. Although the RNAi approach is extremely powerful, concerns still linger about its low efficiency. Here, we developed a CRISPR/Cas9-mediated conditional mutagenesis system by combining tissue-specific expression of Cas9 driven by the Gal4/upstream activating site system with various ubiquitously expressed guide RNA transgenes to effectively inactivate gene expression in a temporally and spatially controlled manner. Furthermore, by including multiple guide RNAs in a transgenic vector to target a single gene, we achieved a high degree of gene mutagenesis in specific tissues. The CRISPR/Cas9-mediated conditional mutagenesis system provides a simple and effective tool for gene function analysis, and complements the existing RNAi approach.
DOI: 10.1371/journal.pbio.1001434
发表时间: 2012
期刊: PLoS biology
影响因子: 9.8
作者:
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