Functional Genomic Analysis of Cell Division by Endoribonuclease-Prepared siRNAs

Abstract

The emergence of RNA interference (RNAi) technology has revolutionized functionalgenomic analyses in cell biology, including the study of cell division. In particular, theintroduction of short interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) hasfacilitated loss-of-function studies in mammalian cell lines. We have pioneered the rapidand cost-efficient generation of libraries of endoribonuclease-prepared short interferingRNAs (esiRNAs) for large-scale genetic screens in mammalian tissue culture cells. Ourfirst pilot screen of 5305 genes in human HeLa cells identified novel genes required forcell division, establishing esiRNA as a sophisticated method for loss-of-function screens.Future genome-wide studies with esiRNA, using image-based assays and videomicroscopy, will help provide novel insights on the molecular mechanisms that governcell division in mammalian cells.