Identification and characterization of essential genes in the human genome.

Abstract

Large-scale genetic analysis of lethal phenotypes has elucidated the molecular underpinnings of many biological processes. Using the bacterial clustered regularly interspaced short palindromic repeats (CRISPR) system, we constructed a genome-wide single-guide RNA library to screen for genes required for proliferation and survival in a human cancer cell line. Our screen revealed the set of cell-essential genes, which was validated with an orthogonal gene-trap-based screen and comparison with yeast gene knockouts. This set is enriched for genes that encode components of fundamental pathways, are expressed at high levels, and contain few inactivating polymorphisms in the human population. We also uncovered a large group of uncharacterized genes involved in RNA processing, a number of whose products localize to the nucleolus. Last, screens in additional cell lines showed a high degree of overlap in gene essentiality but also revealed differences specific to each cell line and cancer type that reflect the developmental origin, oncogenic drivers, paralogous gene expression pattern, and chromosomal structure of each line. These results demonstrate the power of CRISPR-based screens and suggest a general strategy for identifying liabilities in cancer cells.

• Authors: Tim Wang, Kıvanç Birsoy, Nicholas W Hughes, Kevin M Krupczak, Yorick Post, Jenny J Wei, Eric S Lander, David M Sabatini
• Journal: Science (New York, N.Y.) (Science) Vol. 350 Issue 6264 Pg. 1096-101 (Nov 27 2015) ISSN: 1095-9203 [Electronic] United States
• PMID: 26472758 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)
• Copyright: Copyright © 2015, American Association for the Advancement of Science.

Chemical References

• RNA, Guide

Topics

• CRISPR-Cas Systems
• Cell Line, Tumor
• Cell Nucleolus (metabolism)
• Gene Expression Profiling
• Gene Knockout Techniques
• Genes, Essential
• Genetic Testing (methods)
• Genome, Human (genetics)
• Genomic Library
• Humans
• Neoplasms (genetics)
• RNA Processing, Post-Transcriptional (genetics)
• RNA, Guide, Kinetoplastida (genetics)