%0 Journal Article %1 Qi2013Repurposing %A Qi, Lei S. %A Larson, Matthew H. %A Gilbert, Luke A. %A Doudna, Jennifer A. %A Weissman, Jonathan S. %A Arkin, Adam P. %A Lim, Wendell A. %D 2013 %I Cell Press %J Cell %K bt3240 crispr gene-expression %N 5 %P 1173--1183 %R 10.1016/j.cell.2013.02.022 %T Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. %U http://dx.doi.org/10.1016/j.cell.2013.02.022 %V 152 %X Targeted gene regulation on a genome-wide scale is a powerful strategy for interrogating, perturbing, and engineering cellular systems. Here, we develop a method for controlling gene expression based on Cas9, an RNA-guided DNA endonuclease from a type II CRISPR system. We show that a catalytically dead Cas9 lacking endonuclease activity, when coexpressed with a guide RNA, generates a DNA recognition complex that can specifically interfere with transcriptional elongation, RNA polymerase binding, or transcription factor binding. This system, which we call CRISPR interference (CRISPRi), can efficiently repress expression of targeted genes in Escherichia coli, with no detectable off-target effects. CRISPRi can be used to repress multiple target genes simultaneously, and its effects are reversible. We also show evidence that the system can be adapted for gene repression in mammalian cells. This RNA-guided DNA recognition platform provides a simple approach for selectively perturbing gene expression on a genome-wide scale. Copyright \copyright 2013 Elsevier Inc. All rights reserved.

@article{Qi2013Repurposing, abstract = {Targeted gene regulation on a genome-wide scale is a powerful strategy for interrogating, perturbing, and engineering cellular systems. Here, we develop a method for controlling gene expression based on Cas9, an {RNA}-guided {DNA} endonuclease from a type {II} {CRISPR} system. We show that a catalytically dead Cas9 lacking endonuclease activity, when coexpressed with a guide {RNA}, generates a {DNA} recognition complex that can specifically interfere with transcriptional elongation, {RNA} polymerase binding, or transcription factor binding. This system, which we call {CRISPR} interference ({CRISPRi}), can efficiently repress expression of targeted genes in Escherichia coli, with no detectable off-target effects. {CRISPRi} can be used to repress multiple target genes simultaneously, and its effects are reversible. We also show evidence that the system can be adapted for gene repression in mammalian cells. This {RNA}-guided {DNA} recognition platform provides a simple approach for selectively perturbing gene expression on a genome-wide scale. Copyright {\copyright} 2013 Elsevier Inc. All rights reserved.}, added-at = {2018-12-02T16:09:07.000+0100}, author = {Qi, Lei S. and Larson, Matthew H. and Gilbert, Luke A. and Doudna, Jennifer A. and Weissman, Jonathan S. and Arkin, Adam P. and Lim, Wendell A.}, biburl = {https://www.bibsonomy.org/bibtex/2ff3188e37183099f340c9f0c6876d8fb/karthikraman}, citeulike-article-id = {12092414}, citeulike-linkout-0 = {http://www.cell.com//abstract/S0092-8674(13)00211-0}, citeulike-linkout-1 = {http://dx.doi.org/10.1016/j.cell.2013.02.022}, citeulike-linkout-2 = {http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3664290/}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/23452860}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=23452860}, day = 28, doi = {10.1016/j.cell.2013.02.022}, interhash = {57f846e3c131608197161cad37b58656}, intrahash = {ff3188e37183099f340c9f0c6876d8fb}, issn = {1097-4172}, journal = {Cell}, keywords = {bt3240 crispr gene-expression}, month = feb, number = 5, pages = {1173--1183}, pmcid = {PMC3664290}, pmid = {23452860}, posted-at = {2013-03-26 06:01:08}, priority = {2}, publisher = {Cell Press}, timestamp = {2018-12-02T16:09:07.000+0100}, title = {Repurposing {CRISPR} as an {RNA}-guided platform for sequence-specific control of gene expression.}, url = {http://dx.doi.org/10.1016/j.cell.2013.02.022}, volume = 152, year = 2013 }