Identification and characterization of essential genes in the human genome

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...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 2015-11, Vol.350 (6264), p.1096-1101
Main Authors: Wang, Tim, Birsoy, Kivanç, Hughes, Nicholas W., Krupczak, Kevin M., Post, Yorick, Wei, Jenny J., Lander, Eric S., Sabatini, David M.
Format: Article
Language:English
Subjects:
Bacteria
Biotechnology
Cancer
Cell Line, Tumor
Cell Nucleolus - metabolism
CRISPR-Cas Systems
Gene Expression Profiling
Gene Knockout Techniques
Genes
Genes, Essential
Genetic Testing - methods
Genetics
Genome, Human - genetics
Genomes
Genomic Library
Human Genome Project
Humans
Neoplasms - genetics
Ribonucleic acids
RNA Processing, Post-Transcriptional - genetics
RNA, Guide, CRISPR-Cas Systems - genetics
Screens
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Summary: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.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.aac7041