Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Author Luke Gilbert, Max Horlbeck, Britt Adamson, Jacqueline Villalta, Yuwen Chen, Evan Whitehead, Carla Guimaraes, Barbara Panning, Hidde Ploegh, Michael Bassik, Lei Qi, Martin Kampmann, Jonathan Weissman Publication Year 2014 Type Journal Article Abstract While the catalog of mammalian transcripts and their expression levels in different cell types and disease states is rapidly expanding, our understanding of transcript function lags behind. We present a robust technology enabling systematic investigation of the cellular consequences of repressing or inducing individual transcripts. We identify rules for specific targeting of transcriptional repressors (CRISPRi), typically achieving 90%-99% knockdown with minimal off-target effects, and activators (CRISPRa) to endogenous genes via endonuclease-deficient Cas9. Together they enable modulation of gene expression over a ∼1,000-fold range. Using these rules, we construct genome-scale CRISPRi and CRISPRa libraries, each of which we validate with two pooled screens. Growth-based screens identify essential genes, tumor suppressors, and regulators of differentiation. Screens for sensitivity to a cholera-diphtheria toxin provide broad insights into the mechanisms of pathogen entry, retrotranslocation and toxicity. Our results establish CRISPRi and CRISPRa as powerful tools that provide rich and complementary information for mapping complex pathways. Keywords Transcription, Genetic, Humans, Cell Line, Genetic Techniques, Genome, Human, CRISPR-Cas Systems, Cholera Toxin, Diphtheria Toxin Journal Cell Volume 159 Issue 3 Pages 647-61 Date Published 2014 Oct 23 ISSN Number 1097-4172 DOI 10.1016/j.cell.2014.09.029 Alternate Journal Cell PMCID PMC4253859 PMID 25307932 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML