@article {1180686,
	title = {Optimized strategy for in vivo Cas9-activation in Drosophila},
	journal = {Proc Natl Acad Sci U S A},
	volume = {114},
	number = {35},
	year = {2017},
	month = {2017 Aug 29},
	pages = {9409-9414},
	abstract = {While several large-scale resources are available for in vivo loss-of-function studies in Drosophila, an analogous resource for overexpressing genes from their endogenous loci does not exist. We describe a strategy for generating such a resource using Cas9 transcriptional activators (CRISPRa). First, we compare a panel of CRISPRa approaches and demonstrate that, for in vivo studies, dCas9-VPR is the most optimal activator. Next, we demonstrate that this approach is scalable and has a high success rate, as \&gt;75\% of the lines tested activate their target gene. We show that CRISPRa leads to physiologically relevant levels of target gene expression capable of generating strong gain-of-function (GOF) phenotypes in multiple tissues and thus serves as a useful platform for genetic screening. Based on the success of this CRISRPa approach, we are generating a genome-wide collection of flies expressing single-guide RNAs (sgRNAs) for CRISPRa. We also present a collection of more than 30 Gal4 \&gt; UAS:dCas9-VPR lines to aid in using these sgRNA lines for GOF studies in vivo.},
	issn = {1091-6490},
	doi = {10.1073/pnas.1707635114},
	author = {Ewen-Campen, Ben and Yang-Zhou, Donghui and Fernandes, Vit{\'o}ria R and Gonz{\'a}lez, Delfina P and Liu, Lu-Ping and Tao, Rong and Ren, Xingjie and Sun, Jin and Hu, Yanhui and Zirin, Jonathan and Mohr, Stephanie E and Ni, Jian-Quan and Perrimon, Norbert}
}
