Expanding applications of CRISPR/Cas9 technology in Drosophila　理化学研究所多細胞システム形成研究センター（CDB）

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Category	その他
Date and Time	2014-07-17 13:00 - 14:00
Venue	Seminar Room A6F
Speaker	Shu Kondo
Affiliation	Invetebrate Genetics Laboratory,National Institute of Genetics
Title	Expanding applications of CRISPR/Cas9 technology in Drosophila
Host	Erina Kuranaga
Summary	Genome engineering by synthetic nucleases is revolutionizing the way genetic research is done. CRISPR/Cas9 is a recently developed synthetic nuclease that can be programmed to target a new sequence by simply altering a 20-bp sequence in its guide RNA (gRNA) subunit. Here we report three major applications of CRISPR/Cas9 technology in Drosophila. First, we show that transgenic expression of Cas9 protein and gRNA in the germ line induces heritable mutations in target genes in an average of 70% of the F1 progeny. Second, we show that targeted transgene integration, or gene knock-in, can be efficiently induced by co-injection of a targeting vector and gRNA into Cas9-expressing embryos. Finally, we show that tissue-specific expression of Cas9 using the GAL4/UAS system combined with ubiquitous gRNA expression allows spatiotemporally controlled gene inactivation in vivo.