Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells

Author:

Shalem Ophir12,Sanjana Neville E.12,Hartenian Ella1,Shi Xi13,Scott David A.12,Mikkelsen Tarjei S.1,Heckl Dirk4,Ebert Benjamin L.4,Root David E.1,Doench John G.1,Zhang Feng12

Affiliation:

1. Broad Institute of MIT and Harvard, 7 Cambridge Center, Cambridge, MA 02142, USA.

2. McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

3. Stanley Center for Psychiatric Research, Broad Institute of Harvard and MIT, 7 Cambridge Center, Cambridge, MA 02142, USA.

4. Division of Hematology, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA.

Abstract

Improving Whole-Genome Screens Improved methods are needed for the knockout of individual genes in genome-scale functional screens. Wang et al. (p. 80 , published online 12 December) and Shalem et al. (p. 84 , published online 12 December) used the bacterial CRISPR/Cas9 system to power-screen protocols that avoid several of the pitfalls associated with small interfering RNA (siRNA) screens. Genome editing by these methods completely disrupts target genes, thus avoiding weak signals that can occur when transcript abundance is partially decreased by siRNA. Furthermore, gene targeting by the CRISPR system is more precise and appears to produce substantially fewer off-target effects than existing methods.

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

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