Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation
Luke A. Gilbert,Max A. Horlbeck,Britt Adamson,Jacqueline E. Villalta,Yuwen Chen,Evan H. Whitehead,Carla P. Guimaraes,Barbara Panning,Hidde L. Ploegh,Michael C. Bassik,Lei S. Qi,Martin Kampmann,Jonathan S. Weissman +12 more
TLDR
This work identifies rules for specific targeting of transcriptional repressors (CRISPRi), typically achieving 90%-99% knockdown with minimal off-target effects, and activators to endogenous genes via endonuclease-deficient Cas9, which enable modulation of gene expression over a ∼1,000-fold range.About:
This article is published in Cell.The article was published on 2014-10-23 and is currently open access. It has received 2041 citations till now. The article focuses on the topics: CRISPR interference & Cas9.read more
Citations
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Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9
John G. Doench,Nicolo Fusi,Meagan E Sullender,Mudra Hegde,Emma W Vaimberg,Katherine F Donovan,Ian Smith,Zuzana Tothova,Zuzana Tothova,Craig B. Wilen,Robert C. Orchard,Herbert W. Virgin,Jennifer Listgarten,David E. Root +13 more
TL;DR: Recently devised sgRNA design rules are used to create human and mouse genome-wide libraries, perform positive and negative selection screens and observe that the use of these rules produced improved results, and a metric to predict off-target sites is developed.
Journal ArticleDOI
Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex
Silvana Konermann,Mark D. Brigham,Alexandro E. Trevino,Julia Joung,Omar O. Abudayyeh,Clea Barcena,Patrick D. Hsu,Naomi Habib,Jonathan S. Gootenberg,Hiroshi Nishimasu,Osamu Nureki,Feng Zhang +11 more
TL;DR: Structural-guided engineering of a CRISPR-Cas9 complex to mediate efficient transcriptional activation at endogenous genomic loci is described and the potential of Cas9-based activators as a powerful genetic perturbation technology is demonstrated.
Journal ArticleDOI
Crystal structure of Cas9 in complex with guide RNA and target DNA
Takashi Yamano,Hiroshi Nishimasu,Hiroshi Nishimasu,Bernd Zetsche,Hisato Hirano,Ian Slaymaker,Yinqing Li,Iana Fedorova,Takanori Nakane,Kira S. Makarova,Eugene V. Koonin,Ryuichiro Ishitani,Feng Zhang,Osamu Nureki +13 more
TL;DR: In this article, the crystal structure of the CRISPR-associated endonuclease Cas9 in complex with sgRNA and its target DNA at 2.5-A resolution was reported.
Journal ArticleDOI
Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers
Isaac B. Hilton,Anthony D'Ippolito,Christopher M. Vockley,Pratiksha I. Thakore,Gregory E. Crawford,Timothy E. Reddy,Charles A. Gersbach +6 more
TL;DR: A programmable, CRISPR-Cas9-based acetyltransferase consisting of the nuclease-null dCas9 protein fused to the catalytic core of the human acetyl transferase p300 is described, leading to robust transcriptional activation of target genes from promoters and both proximal and distal enhancers.
Journal ArticleDOI
High-throughput functional genomics using CRISPR–Cas9
TL;DR: A review of the latest applications of CRISPR-Cas9 in mammalian functional genomics screens is presented in this article, which covers related genome-scale applications of Cas9 for either gene knockout or transcriptional modulation.
References
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