Cell-Based RNAi Assay Development for HTS RNAi Loss-of-Function Screens

Abstract

RNA interference (RNAi) experiments are powerful tools to study regulation of gene expression to understand target validation in drug discovery. RNAi screens can be run in a loss-of-function or a synthetic lethality mode using small synthetic RNAs (siRNA or shRNA), specific for genes of interest transfected into cells. In this chapter, the basic protocols in the design and development on RNAi screens, assay validation and optimization, data analysis and hit selection criteria in typical HTS applications are described.