ShRNA screen identifies novel mediator of PDAC metastasis in vivo

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. 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.

本研究开展了一项短发夹RNA（short hairpin RNA, shRNA）筛选实验，旨在探究胰腺癌相关基因在体内转移性生长中的作用。研究共筛选得到284个候选基因，筛选依据包括基因与胰腺癌的相关性、胰腺癌中的突变频率，或其受Ras调控的特性。分别在注射前的细胞（第0天）、分离得到的肺肿瘤组织（第28天）以及体外培养的细胞（第28天）中，对shRNA的丰度进行了检测。每个靶基因对应3~4条shRNA，最终构建的shRNA文库共包含1013条shRNA。共生成19个shRNA混合池，每个混合池均包含8条对照shRNA，用于评估不同混合池间的实验一致性。