Rapid modeling of cooperating genetic events in cancer through somatic genome editing

Cancer is a multistep process that involves mutations and other alterations in oncogenes and tumor suppressor genes. Genome sequencing studies have identified a large collection of genetic alterations that occur in human cancers. However, the determination of which mutations are causally related to tumorigenesis remains a major challenge for cancer genome sequencing. Here we describe a novel CRISPR/Cas9-based approach for rapid functional investigation of candidate genes in well-established autochthonous mouse models of cancer. Using a KrasG12D-driven lung cancer model5, we performed functional characterization of a panel of tumor suppressor genes with known loss-of-function alterations in human lung cancer. Cre-dependent somatic activation of oncogenic KrasG12D combined with CRISPR/Cas9-mediated genome editing of tumor suppressor genes resulted in lung adenocarcinomas with distinct histopathological and molecular features. This rapid somatic genome engineering approach enables functional characterization of putative driver genetic events that cooperate with oncogenic Kras in the context of an autochthonous mouse model. We anticipate that this approach can be used to systematically dissect the complex catalog of mutations identified in cancer genome sequencing studies.