| Speaker: |
Shu Kondo
Invetebrate Genetics Laboratory,National Institute of Genetics |
| Title: |
Expanding applications of CRISPR/Cas9 technology in Drosophila |
| 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. |
