Versatile in vivo regulation of tumor phenotypes by dCas9-mediated transcriptional perturbation
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Author(s)
Horlbeck, Max A.
Gilbert, Luke A.
Weissman, Jonathan S.
Bruno, Peter Michael
Hemann, Michael
Braun, Christian Joerg
Date Issued
June 2016
Journal
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences (U.S.)
Citation
Braun, Christian J. et al. “Versatile in Vivo Regulation of Tumor Phenotypes by dCas9-Mediated Transcriptional Perturbation.” Proceedings of the National Academy of Sciences 113, 27 (June 2016): E3892–E3900 © 2016 The Authors
Version
Final published version
Abstract
Targeted transcriptional regulation is a powerful tool to study genetic mediators of cellular behavior. Here, we show that catalytically dead Cas9 (dCas9) targeted to genomic regions upstream or downstream of the transcription start site allows for specific and sustainable gene-expression level alterations in tumor cells in vitro and in syngeneic immune-competent mouse models. We used this approach for a high-coverage pooled gene-activation screen in vivo and discovered previously unidentified modulators of tumor growth and therapeutic response. Moreover, by using dCas9 linked to an activation domain, we can either enhance or suppress target gene expression simply by changing the genetic location of dCas9 binding relative to the transcription start site. We demonstrate that these directed changes in gene-transcription levels occur with minimal off-target effects. Our findings highlight the use of dCas9-mediated transcriptional regulation as a versatile tool to reproducibly interrogate tumor phenotypes in vivo. Keywords: cancer genetics; cancer models; cancer therapeutic resistance; gene regulation; CRISPR
MIT Department
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1073/PNAS.1600582113
