RNA Interference (RNAi) Screening in

In the last decade, RNA interference (RNAi), a cellular mechanism that uses RNA-guided degradation of messenger RNA transcripts, has had an important impact on identifying and characterizing gene function. First discovered in , RNAi can be used to silence the expression of genes through introduction of exogenous double-stranded RNA into cells. In , RNAi has been applied in cultured cells or to perturb the function of single genes or to systematically probe gene function on a genome-wide scale. In this review, we will describe the use of RNAi to study gene function in with a particular focus on high-throughput screening methods applied in cultured cells. We will discuss available reagent libraries and cell lines, methodological approaches for cell-based assays, and computational methods for the analysis of high-throughput screens. Furthermore, we will review the generation and use of genome-scale RNAi libraries for tissue-specific knockdown analysis and discuss the differences and similarities with the use of genome-engineering methods such as CRISPR/Cas9 for functional analysis.