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dc.contributor.authorEwen-Campen, Benjamin
dc.contributor.authorYang-Zhou, Donghui
dc.contributor.authorFernandes, Vitória R.
dc.contributor.authorGonzález, Delfina P.
dc.contributor.authorLiu, Lu-Ping
dc.contributor.authorTao, Rong
dc.contributor.authorRen, Xingjie
dc.contributor.authorSun, Jin
dc.contributor.authorHu, Yanhui
dc.contributor.authorZirin, Jonathan
dc.contributor.authorMohr, Stephanie
dc.contributor.authorNi, Jian-Quan
dc.contributor.authorPerrimon, Norbert
dc.date.accessioned2021-09-29T15:40:54Z
dc.date.issued2017-08-29
dc.identifier.citationEwen-Campen, Benjamin, Donghui Yang-Zhou, Vitória R. Fernandes, Delfina P. González, Lu-Ping Liu, Rong Tao, Xingjie Ren et al. "Optimized Strategy for in Vivo Cas9-Activation in Drosophila." Proceedings of the National Academy of Sciences 114, no. 35 (2017): 9409-9414. DOI: 10.1073/pnas.1707635114
dc.identifier.issn0027-8424en_US
dc.identifier.issn1091-6490en_US
dc.identifier.urihttps://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37369614*
dc.description.abstractWhile 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 >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 > UAS:dCas9-VPR lines to aid in using these sgRNA lines for GOF studies in vivo.en_US
dc.language.isoen_USen_US
dc.publisherNational Academy of Sciencesen_US
dc.relation.isversionofdoi:10.1073/pnas.1707635114en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC5584449/en_US
dash.licenseMETA_ONLY
dc.subjectResearch Subject Categories::NATURAL SCIENCES::Biology::Other biology::Functional genomicsen_US
dc.titleOptimized Strategy for in Vivo Cas9-Activation in Drosophilaen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalProceedings of the National Academy of Sciencesen_US
dash.depositing.authorPerrimon, Norbert
dc.date.available2021-09-29T15:40:54Z
dc.identifier.doi10.1073/pnas.1707635114
dash.source.volume114en_US
dash.source.page9409-9414en_US
dash.source.issue35en_US
dash.contributor.affiliatedMohr, Stephanie
dash.contributor.affiliatedEwen-Campen, Benjamin
dash.contributor.affiliatedTao, Rong
dash.contributor.affiliatedHu, Yanhui
dash.contributor.affiliatedZirin, Jonathan
dash.contributor.affiliatedPerrimon, Norbert


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