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Next generation CRISPR/Cas9 transcriptional activation in Drosophila using flySAM

By Yu Jia, Rong-Gang Xu, Xingjie Ren, Ben Ewen-Campen, Rajendhran Rajakumar, Jonathan Zirin, Donghui Yang-Zhou, Ruibao Zhu, Fang Wang, Decai Mao, Ping Peng, Huan-Huan Qiao, Xia Wang, Lu-Ping Liu, Bowen Xu, Jun-Yuan Ji, Qingfei Liu, Jin Sun, Norbert Perrimon, Jian-Quan Ni

Posted 31 Jan 2018
bioRxiv DOI: 10.1101/252031 (published DOI: 10.1073/pnas.1800677115)

CRISPR/Cas9-based transcriptional activation (CRISPRa) has recently emerged as a powerful and scalable technique for systematic over-expression genetic analysis in Drosophila melanogaster. We present flySAM, a potent new tool for in vivo CRISPRa, which offers a major improvement over existing strategies in terms of effectiveness, scalability, and ease-of-use. flySAM outperforms existing in vivo CRISPRa strategies, and approximates phenotypes obtained using traditional Gal4-UAS over-expression. Further, because flySAM typically only requires a single sgRNA, it dramatically improves scalability. We use flySAM to demonstrate multiplexed CRISPRa, which has not been previously shown in vivo. In addition, we have simplified the experimental usage of flySAM by creating a single vector encoding both the UAS:Cas9-activator and the sgRNA, allowing for inducible CRISPRa in a single genetic cross. flySAM will thus replace previous CRISPRa strategies as the basis of our growing genome-wide transgenic over-expression resource, TRiP-OE.

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