Simple Gene Knockdown in Endothelial Cells Using Short Interfering RNA Oligonucleotides

Zarah B Tabrizi; Kathryn R Green; Amy P Lynch; Nada S Ahmed; Nicholas Beazley-Long; Andrew V Benest

doi:10.1007/978-1-0716-2059-5_19

Simple Gene Knockdown in Endothelial Cells Using Short Interfering RNA Oligonucleotides

Methods Mol Biol. 2022:2441:251-255. doi: 10.1007/978-1-0716-2059-5_19.

Authors

Zarah B Tabrizi¹, Kathryn R Green¹, Amy P Lynch¹, Nada S Ahmed¹, Nicholas Beazley-Long¹, Andrew V Benest²

Affiliations

¹ Endothelial Quiescence Group, Centre for Cancer Sciences, Biodiscovery Institute, School of Medicine, University of Nottingham, Nottingham, UK.
² Endothelial Quiescence Group, Centre for Cancer Sciences, Biodiscovery Institute, School of Medicine, University of Nottingham, Nottingham, UK. Andrew.Benest@nottingham.ac.uk.

PMID: 35099742
DOI: 10.1007/978-1-0716-2059-5_19

Abstract

Cell transfection using short interfering RNAs (siRNAs) is a widely used technique to perform loss of function studies by "knocking down" genes of interest. Oftentimes, primary cells can be difficult to transfect, but here we provide a simple and robust method using cultured endothelial cells and routine transfection reagents. Knockdown studies can be used to complement overexpression studies and validate biochemical pathway analysis, as well as functional assays. The enclosed protocol will compliment other in vitro assays detailed in this edition.

Keywords: HDLEC; HUVEC; Knockdown; RNA interference; Transfection; siRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Endothelial Cells* / metabolism
Gene Knockdown Techniques
Oligonucleotides* / metabolism
RNA Interference
RNA, Small Interfering / genetics
RNA, Small Interfering / metabolism
Transfection

Substances

Oligonucleotides
RNA, Small Interfering

Grants and funding

PG/18/31/33759/BHF_/British Heart Foundation/United Kingdom