RNA Interference-Mediated Inhibition of ESCRT in Mammalian Cells

Katherine Bowers

doi:10.1007/978-1-4939-9492-2_22

RNA Interference-Mediated Inhibition of ESCRT in Mammalian Cells

Methods Mol Biol. 2019:1998:305-318. doi: 10.1007/978-1-4939-9492-2_22.

Author

Katherine Bowers¹

Affiliation

¹ Division of Biosciences, Institute of Structural and Molecular Biology, University College London, Gower Street, London, WC1E 6BT, UK. katherine.bowers@ucl.ac.uk.

PMID: 31250311
DOI: 10.1007/978-1-4939-9492-2_22

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).

Keywords: ESCRT; Endosomes; Protein depletion; RNAi; siRNA.

MeSH terms

Cell Culture Techniques / methods
Endosomal Sorting Complexes Required for Transport / genetics*
Endosomal Sorting Complexes Required for Transport / metabolism
Gene Knockdown Techniques / methods*
HeLa Cells
Humans
RNA Interference*
RNA, Small Interfering / genetics
Transfection / methods

Substances

Endosomal Sorting Complexes Required for Transport
RNA, Small Interfering