Synthetic-Lethality Screening For Critical Genes Involved In Drug Resistance Of Pancreatic Cancer

Objevtive The goals of this study is to apply genome-wide synthetic lethality screening for the discovery of the critical genes invovled in therapeutic drug resistance of Pancreatic cancer; to provide novel biomarkers, therapeutical targets and novel combination therapeutics for Pancreatic cancer.Methods We constructed piggyBac transposon based genome-wide screening vectors by molecular cloning; characterized human Pancreatic cancer cell line (Panc-1); generated stable Panc-1cell lines with Tetracycline-mediated gene regulation with DNA transfection and Luciferase based reporter assay screening; established genome-wide random mutagenesis libraries of Panc-1cells by large scale electroporation and piggyBac mediated genetic recombination; and further carried out preliminary screening of synthetic lethality with Gemcitabine.Results We constructed piggyBac transposon based genome-wide screening vectors; generated highly inducible Panc-1cell lines with Tetracycline-mediated gene regulation, i.e. Panc-1-9.3and Panc-1-17B1; and successfully established genome-wide random mutagenesis libraries of Panc-1cells, that will be applied to screening of synthetic lethality with Gemcitabine.Conclusion We generated highly inducible tetracycline regulated human pancreatic caner cell lines, and random genome-wide mutagenesis libraries. This provides a valuable platform for the genetic screening of synthetic lethality with chemotherapeutics in pancreatic cancer.