RNA Interference-Mediated Inhibition of ESCRT in Mammalian Cells

Abstract

Specific depletion of proteins from cultured cells using RNA interference (RNAi) has been a useful technique in assessing protein function for many years. RNAi allows the degradation of specific, targeted mRNA, allowing the effects of protein depletion on cellular processes to be examined. Here, I present a protocol for the depletion of proteins from cultured HeLa cells and list specific reagents and considerations for targeting the endosomal sorting complexes required for transport (ESCRT).