You are here

Nat Biotechnol DOI:10.1038/nbt.2951

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

Publication TypeJournal Article
Year of Publication2014
AuthorsHeckl, D, Kowalczyk, MS, Yudovich, D, Belizaire, R, Puram, RV, McConkey, ME, Thielke, A, Aster, JC, Regev, A, Ebert, BL
JournalNat Biotechnol
Date Published2014 Sep
KeywordsAnimals, Bone Marrow Neoplasms, Clustered Regularly Interspaced Short Palindromic Repeats, Disease Models, Animal, Mice

Genome sequencing studies have shown that human malignancies often bear mutations in four or more driver genes, 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 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.


Alternate JournalNat. Biotechnol.
PubMed ID24952903
PubMed Central IDPMC4160386
Grant ListR01 HL082945 / HL / NHLBI NIH HHS / United States
T32 GM007753 / GM / NIGMS NIH HHS / United States
P01 CA066996 / CA / NCI NIH HHS / United States
P01 CA108631 / CA / NCI NIH HHS / United States
P50 HG006193 / HG / NHGRI NIH HHS / United States
5P50HG006193-02 / HG / NHGRI NIH HHS / United States