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Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing

2014; Nature Portfolio; Volume: 32; Issue: 9 Linguagem: Inglês

10.1038/nbt.2951

1546-1696

Dirk Heckl, Monika S. Kowalczyk, David Yudovich, Roger Belizaire, Rishi V. Puram, Marie McConkey, Anne L. Thielke, Jon C. Aster, Aviv Regev, Benjamin L. Ebert,

Insect Resistance and Genetics

The genetic complexity of human leukemia is modeled using the CRISPR-Cas9 system in mice. Genome sequencing studies have shown that human malignancies often bear mutations in four or more driver genes1, 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 editing2,3,4 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.