Using RNA interference to study protein function.
RNA interference can be extremely useful in determining the function of an endogenously-expressed protein in its normal cellular environment. In this chapter, we describe a method that uses small interfering RNA (siRNA) to knock down mRNA and protein expression in cultured cells so that the effect of a putative regulatory protein on gene expression can be delineated. Methods of assessing the effectiveness of the siRNA procedure using real time quantitative PCR and Western analysis are also included.
Curtis
Carol D
CD
Department of Molecular and Integrative Physiology, University of Illinois, Urbana, IL, USA.
Nardulli
Ann M
AM
D045744:Cell Line, Tumor; D002478:Cells, Cultured; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods
D011506:Proteins; D034741:RNA, Small Interfering