Modeling Lung Adenocarcinoma Gene Fusions In Vivo With CRISPR-Cas12a

Lung adenocarcinoma (LUAD) is the most common subtype of non-small cell lung cancer. 10% of all LUAD cases are driven by gene fusions, where one gene partner fuses to an oncogenic kinase, promoting kinase expression, activity, and thus dysregulating downstream signaling pathways. Although prior studies that have modeled EML4-ALK translocations have used Cas9 and Cas12a in mice, in vivo models of LUAD fusions remain limited. To facilitate research on fusion-driven tumorigenesis, this study attempts to induce 19 LUAD gene fusions (spanning eight different kinases) and 15 ROS1 fusions in lung epithelial cells in Cas12a-expressing mice. In these experiments, 3-guide crRNA arrays were designed with one guide targeting each intronic fusion breakpoint and a third guide for Tp53 inactivation. 3-guide arrays were subsequently cloned into barcoded lentiviral vectors and lentivirus was delivered to the lungs of Cas12a-expressing mice. Overall, this study seeks to provide models to further investigate fusion-targeting drug therapies, fusion mechanisms, and fusion-activated signaling pathways.