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 (sgRNA) library to screen for genes...
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| Format: | Article |
| Language: | en_US |
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American Association for the Advancement of Science (AAAS)
2017
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| Online Access: | http://hdl.handle.net/1721.1/106204 https://orcid.org/0000-0002-4227-5163 https://orcid.org/0000-0002-1446-7256 |
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| author | Hughes, N. W. Wang, Tim Birsoy, Kivanc Krupczak, Kevin M. Post, Yorick Lander, Eric Steven Sabatini, David Wei, Jenny J. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Hughes, N. W. Wang, Tim Birsoy, Kivanc Krupczak, Kevin M. Post, Yorick Lander, Eric Steven Sabatini, David Wei, Jenny J. |
| author_sort | Hughes, N. W. |
| collection | MIT |
| description | 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 (sgRNA) 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 by 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. Lastly,
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. |
| first_indexed | 2024-09-23T12:53:44Z |
| format | Article |
| id | mit-1721.1/106204 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T12:53:44Z |
| publishDate | 2017 |
| publisher | American Association for the Advancement of Science (AAAS) |
| record_format | dspace |
| spelling | mit-1721.1/1062042022-10-01T11:46:25Z Identification and characterization of essential genes in the human genome Hughes, N. W. Wang, Tim Birsoy, Kivanc Krupczak, Kevin M. Post, Yorick Lander, Eric Steven Sabatini, David Wei, Jenny J. Massachusetts Institute of Technology. Department of Biology Whitehead Institute for Biomedical Research Koch Institute for Integrative Cancer Research at MIT Wang, Tim Birsoy, Kivanc Krupczak, Kevin M. Post, Yorick Wei, Jenny J Lander, Eric Steven Sabatini, David 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 (sgRNA) 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 by 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. Lastly, 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. National Institutes of Health (U.S.) (Grant CA103866) National Human Genome Research Institute (U.S.) (Grant 2U54HG003067-1) National Science Foundation (U.S.) Whitaker Health Sciences Fund 2017-01-05T16:46:34Z 2017-01-05T16:46:34Z 2015-11 2014-10 Article http://purl.org/eprint/type/JournalArticle 0036-8075 1095-9203 http://hdl.handle.net/1721.1/106204 Wang, T. et al. “Identification and Characterization of Essential Genes in the Human Genome.” Science 350.6264 (2015): 1096–1101. https://orcid.org/0000-0002-4227-5163 https://orcid.org/0000-0002-1446-7256 en_US http://dx.doi.org/10.1126/science.aac7041 Science Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Association for the Advancement of Science (AAAS) PMC |
| spellingShingle | Hughes, N. W. Wang, Tim Birsoy, Kivanc Krupczak, Kevin M. Post, Yorick Lander, Eric Steven Sabatini, David Wei, Jenny J. Identification and characterization of essential genes in the human genome |
| title | Identification and characterization of essential genes in the human genome |
| title_full | Identification and characterization of essential genes in the human genome |
| title_fullStr | Identification and characterization of essential genes in the human genome |
| title_full_unstemmed | Identification and characterization of essential genes in the human genome |
| title_short | Identification and characterization of essential genes in the human genome |
| title_sort | identification and characterization of essential genes in the human genome |
| url | http://hdl.handle.net/1721.1/106204 https://orcid.org/0000-0002-4227-5163 https://orcid.org/0000-0002-1446-7256 |
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