Heckl Dirk, Kowalczyk Monika S, Yudovich David, Belizaire Roger, Puram Rishi V, McConkey Marie E, Thielke Anne, Aster Jon C, Regev Aviv, Ebert Benjamin L
1] Division of Hematology, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. [2].
1] Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA. [2].
Nat Biotechnol. 2014 Sep;32(9):941-6. doi: 10.1038/nbt.2951. Epub 2014 Jun 22.
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.
基因组测序研究表明，人类恶性肿瘤通常携带四个或更多驱动基因的突变，但使用传统育种方法在小鼠模型中重现这种程度的遗传复杂性却很困难。在此，我们利用基因组编辑的CRISPR-Cas9系统来克服这一限制。通过慢病毒载体传递小向导RNA（sgRNA）和Cas9的组合，我们在单个小鼠造血干细胞（HSC）中对多达五个基因进行了修饰，导致克隆性增殖和髓系恶性肿瘤。我们由此生成了急性髓系白血病（AML）模型，这些模型在编码表观遗传修饰因子、转录因子和细胞因子信号传导介质的基因中存在协同突变，重现了在患者中观察到的突变组合。我们的结果表明，慢病毒传递的sgRNA:Cas9基因组编辑对于构建更能反映人类疾病复杂性的广泛体内癌症模型应该是有用的。
