10X single-cell RNA-seq profiling of ccRCC model tumors and normal kidney tissue
To achieve a robust immunocompetent mouse model of human ccRCC, we directly injected adenovirus-associated viruses (AAVs) encoding sgRNAs against VHL and other known/suspected ccRCC tumor suppressor genes into the kidneys of C57BL/6 mice under conditions where Cas9 was under the control of the Pax8 promoter to induce kidney tumors. An AAV targeting Vhl, Pbrm1, Keap1 and Tsc1 reproducibly caused macroscopic ccRCCs that partially resembled human ccRCC tumors with respect to transcriptome and cell of origin and responded to a ccRCC standard of care agent, axitinib. Overall design: LSL-Cas9/+ mice were crossed with Pax8 Cre mice to produce LSL-Cas9/+; Pax8 Cre/+ mice and orthotopically injected in the kidney with AAV expressing sgRNAs for Keap1, Pbrm1, Vhl and Tsc1. Four model tumors and four normal kidney samples were collected from mice, dissociated, and subjected to single cell RNA-sequencing.