siRNA screening of nuclear proteins

Abstract

Functional gene screening is a basic strategy for identifying candidate genes that are responsible for biological processes of interest to researchers. RNA-mediated interference (RNAi)-based screening is one such approach. Although the knockdown technique using siRNA is widely accepted, careful evaluation is required to avoid false positive candidates as much as possible, because various cellular responses can be caused by events other than the target gene disruption. In this chapter, we introduce a method for a relatively small-scale siRNA screening that is capable of fi nding RNA binding proteins that regulate a specifi c RNA that is a structural component of a nuclear compartment.