Genome-wide CRISPR Screen in a Mouse Model of Tumor Growth and Metastasis
Genetic screens are powerful tools for identifying genes responsible for diverse phenotypes. Here we describe a genome-wide CRISPR/Cas9-mediated loss-of-function screen in tumor growth and metastasis. We mutagenized a non-metastatic mouse cancer cell line using a genome-scale library with 67,405 sin...
| Main Authors: | , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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Elsevier
2017
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| Online Access: | http://hdl.handle.net/1721.1/111184 https://orcid.org/0000-0002-2639-9879 https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0003-1465-1691 |
| _version_ | 1826189671213826048 |
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| author | Lee, Kyungheon Song, Jun Weissleder, Ralph Lee, Hakho Chen, Sidi Sanjana, Neville E Zheng, Kaijie Shalem, Ophir Shi, Xi Scott, David Arthur Pan, Jennifer Q. Zhang, Feng Sharp, Phillip A. |
| author2 | Broad Institute of MIT and Harvard |
| author_facet | Broad Institute of MIT and Harvard Lee, Kyungheon Song, Jun Weissleder, Ralph Lee, Hakho Chen, Sidi Sanjana, Neville E Zheng, Kaijie Shalem, Ophir Shi, Xi Scott, David Arthur Pan, Jennifer Q. Zhang, Feng Sharp, Phillip A. |
| author_sort | Lee, Kyungheon |
| collection | MIT |
| description | Genetic screens are powerful tools for identifying genes responsible for diverse phenotypes. Here we describe a genome-wide CRISPR/Cas9-mediated loss-of-function screen in tumor growth and metastasis. We mutagenized a non-metastatic mouse cancer cell line using a genome-scale library with 67,405 single-guide RNAs (sgRNAs). The mutant cell pool rapidly generates metastases when transplanted into immunocompromised mice. Enriched sgRNAs in lung metastases and late-stage primary tumors were found to target a small set of genes, suggesting that specific loss-of-function mutations drive tumor growth and metastasis. Individual sgRNAs and a small pool of 624 sgRNAs targeting the top-scoring genes from the primary screen dramatically accelerate metastasis. In all of these experiments, the effect of mutations on primary tumor growth positively correlates with the development of metastases. Our study demonstrates Cas9-based screening as a robust method to systematically assay gene phenotypes in cancer evolution in vivo. |
| first_indexed | 2024-09-23T08:19:17Z |
| format | Article |
| id | mit-1721.1/111184 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:19:17Z |
| publishDate | 2017 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1111842022-09-23T12:19:00Z Genome-wide CRISPR Screen in a Mouse Model of Tumor Growth and Metastasis Lee, Kyungheon Song, Jun Weissleder, Ralph Lee, Hakho Chen, Sidi Sanjana, Neville E Zheng, Kaijie Shalem, Ophir Shi, Xi Scott, David Arthur Pan, Jennifer Q. Zhang, Feng Sharp, Phillip A. Broad Institute of MIT and Harvard Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences McGovern Institute for Brain Research at MIT Koch Institute for Integrative Cancer Research at MIT Chen, Sidi Sanjana, Neville E Zheng, Kaijie Shalem, Ophir Shi, Xi Scott, David Arthur Pan, Jennifer Q. Zhang, Feng Sharp, Phillip A. Genetic screens are powerful tools for identifying genes responsible for diverse phenotypes. Here we describe a genome-wide CRISPR/Cas9-mediated loss-of-function screen in tumor growth and metastasis. We mutagenized a non-metastatic mouse cancer cell line using a genome-scale library with 67,405 single-guide RNAs (sgRNAs). The mutant cell pool rapidly generates metastases when transplanted into immunocompromised mice. Enriched sgRNAs in lung metastases and late-stage primary tumors were found to target a small set of genes, suggesting that specific loss-of-function mutations drive tumor growth and metastasis. Individual sgRNAs and a small pool of 624 sgRNAs targeting the top-scoring genes from the primary screen dramatically accelerate metastasis. In all of these experiments, the effect of mutations on primary tumor growth positively correlates with the development of metastases. Our study demonstrates Cas9-based screening as a robust method to systematically assay gene phenotypes in cancer evolution in vivo. National Institutes of Health (U.S.) (Grant R01-CA133404) National Cancer Institute (U.S.) (Grant U54 CA151884) National Cancer Institute (U.S.) (Grant P30-CA14051) National Institute of Mental Health (U.S.) (Grant 5DP1-MH100706) National Institute of Diabetes and Digestive and Kidney Diseases (U.S.) (Grant 5R01-DK097768) 2017-09-13T15:05:14Z 2017-09-13T15:05:14Z 2015-03 2015-02 Article http://purl.org/eprint/type/JournalArticle 0092-8674 1097-4172 http://hdl.handle.net/1721.1/111184 Chen, Sidi et al. “Genome-Wide CRISPR Screen in a Mouse Model of Tumor Growth and Metastasis.” Cell 160, 6 (March 2015): 1246–1260 © 2015 Elsevier Inc https://orcid.org/0000-0002-2639-9879 https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0003-1465-1691 en_US http://dx.doi.org/10.1016/j.cell.2015.02.038 Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Lee, Kyungheon Song, Jun Weissleder, Ralph Lee, Hakho Chen, Sidi Sanjana, Neville E Zheng, Kaijie Shalem, Ophir Shi, Xi Scott, David Arthur Pan, Jennifer Q. Zhang, Feng Sharp, Phillip A. Genome-wide CRISPR Screen in a Mouse Model of Tumor Growth and Metastasis |
| title | Genome-wide CRISPR Screen in a Mouse Model of Tumor Growth and Metastasis |
| title_full | Genome-wide CRISPR Screen in a Mouse Model of Tumor Growth and Metastasis |
| title_fullStr | Genome-wide CRISPR Screen in a Mouse Model of Tumor Growth and Metastasis |
| title_full_unstemmed | Genome-wide CRISPR Screen in a Mouse Model of Tumor Growth and Metastasis |
| title_short | Genome-wide CRISPR Screen in a Mouse Model of Tumor Growth and Metastasis |
| title_sort | genome wide crispr screen in a mouse model of tumor growth and metastasis |
| url | http://hdl.handle.net/1721.1/111184 https://orcid.org/0000-0002-2639-9879 https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0003-1465-1691 |
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