Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing
Genome sequencing studies have shown that human malignancies often bear mutations in four or more driver genes[superscript 1], but it is difficult to recapitulate this degree of genetic complexity in mouse models using conventional breeding. Here we use the CRISPR-Cas9 system of genome editing[super...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2015
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| Online Access: | http://hdl.handle.net/1721.1/96710 https://orcid.org/0000-0001-8567-2049 |
| _version_ | 1811090677146058752 |
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| author | Heckl, Dirk Kowalczyk, Monika S. Yudovich, David Belizaire, Roger Puram, Rishi V. McConkey, Marie Thielke, Anne Aster, Jon C. Regev, Aviv Ebert, Benjamin L. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Heckl, Dirk Kowalczyk, Monika S. Yudovich, David Belizaire, Roger Puram, Rishi V. McConkey, Marie Thielke, Anne Aster, Jon C. Regev, Aviv Ebert, Benjamin L. |
| author_sort | Heckl, Dirk |
| collection | MIT |
| description | Genome sequencing studies have shown that human malignancies often bear mutations in four or more driver genes[superscript 1], but it is difficult to recapitulate this degree of genetic complexity in mouse models using conventional breeding. Here we use the CRISPR-Cas9 system of genome editing[superscript 2, 3, 4] to overcome this limitation. By delivering combinations of small guide RNAs (sgRNAs) and Cas9 with a lentiviral vector, we modified up to five genes in a single mouse hematopoietic stem cell (HSC), leading to clonal outgrowth and myeloid malignancy. We thereby generated models of acute myeloid leukemia (AML) with cooperating mutations in genes encoding epigenetic modifiers, transcription factors and mediators of cytokine signaling, recapitulating the combinations of mutations observed in patients. Our results suggest that lentivirus-delivered sgRNA:Cas9 genome editing should be useful to engineer a broad array of in vivo cancer models that better reflect the complexity of human disease. |
| first_indexed | 2024-09-23T14:50:07Z |
| format | Article |
| id | mit-1721.1/96710 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T14:50:07Z |
| publishDate | 2015 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/967102022-09-29T10:50:43Z Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing Heckl, Dirk Kowalczyk, Monika S. Yudovich, David Belizaire, Roger Puram, Rishi V. McConkey, Marie Thielke, Anne Aster, Jon C. Regev, Aviv Ebert, Benjamin L. Massachusetts Institute of Technology. Department of Biology Regev, Aviv Genome sequencing studies have shown that human malignancies often bear mutations in four or more driver genes[superscript 1], but it is difficult to recapitulate this degree of genetic complexity in mouse models using conventional breeding. Here we use the CRISPR-Cas9 system of genome editing[superscript 2, 3, 4] to overcome this limitation. By delivering combinations of small guide RNAs (sgRNAs) and Cas9 with a lentiviral vector, we modified up to five genes in a single mouse hematopoietic stem cell (HSC), leading to clonal outgrowth and myeloid malignancy. We thereby generated models of acute myeloid leukemia (AML) with cooperating mutations in genes encoding epigenetic modifiers, transcription factors and mediators of cytokine signaling, recapitulating the combinations of mutations observed in patients. Our results suggest that lentivirus-delivered sgRNA:Cas9 genome editing should be useful to engineer a broad array of in vivo cancer models that better reflect the complexity of human disease. National Institutes of Health (U.S.) (P01 CA108631) Leukemia & Lymphoma Society of America (Scholar Award) trategic Pharma-Academic Research Consortium for Translational Medicine (SPARC) National Human Genome Research Institute (U.S.). Centers of Excellence in Genomic Science (Grant 5P50HG006193-02) Broad Institute of MIT and Harvard 2015-04-22T18:42:07Z 2015-04-22T18:42:07Z 2014-06 2014-04 Article http://purl.org/eprint/type/JournalArticle 1087-0156 1546-1696 http://hdl.handle.net/1721.1/96710 Heckl, Dirk, Monika S Kowalczyk, David Yudovich, Roger Belizaire, Rishi V Puram, Marie E McConkey, Anne Thielke, Jon C Aster, Aviv Regev, and Benjamin L Ebert. “Generation of Mouse Models of Myeloid Malignancy with Combinatorial Genetic Lesions Using CRISPR-Cas9 Genome Editing.” Nature Biotechnology 32, no. 9 (June 22, 2014): 941–946. https://orcid.org/0000-0001-8567-2049 en_US http://dx.doi.org/10.1038/nbt.2951 Nature Biotechnology Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Nature Publishing Group PMC |
| spellingShingle | Heckl, Dirk Kowalczyk, Monika S. Yudovich, David Belizaire, Roger Puram, Rishi V. McConkey, Marie Thielke, Anne Aster, Jon C. Regev, Aviv Ebert, Benjamin L. Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing |
| title | Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing |
| title_full | Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing |
| title_fullStr | Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing |
| title_full_unstemmed | Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing |
| title_short | Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing |
| title_sort | generation of mouse models of myeloid malignancy with combinatorial genetic lesions using crispr cas9 genome editing |
| url | http://hdl.handle.net/1721.1/96710 https://orcid.org/0000-0001-8567-2049 |
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