An shRNA screen with the aim to investigate the role of pancreatic cancer-associated genes in the metastatic outgrowth in vivo was carried out. The set of candidate genes (284) was selected based on their association with pancreatic cancer, frequency of mutation in pancreatic cancer or on being Ras-regulated.
Overall design
The abundance of shRNAs was assessed in cells prior to injection (Day 0), in isolated lung tumours (Day 28), and in cells grown in culture (Day 28). Each gene was targeted with 3-4 shRNA producing a library size of 1013 shRNA. Nineteen shRNA pools were generated and each contained 8 control shRNA to assess consistency across pools.