RNAi screening in Trp53-/- mice to identify tumor suppressor genes in liver cancer

To identify mediators of Trp53 independent tumor suppressive functions of Cdkn2aArf in the mouse liver, we set up an in vivo RNAi screen. We compiled a miRNA-based shRNA library targeting upregulated genes in Trp53–/– mouse livers (compared to NrasG12V; Cdkn2aArf–/– livers upon NrasG12V delivery), published p19Arf interaction partners and genes involved in sumoylation. This shRNA library was delivered via transposon-based gene transfer together with NrasG12V into Trp53–/– livers. Seven weeks later we assessed the tumor burden in mice injected with different shRNA library pools via Illumina deep sequencing.