Small interfering RNA-mediated connexin gene knockdown in vascular endothelial and smooth muscle cells

Good, M. E., Begandt, D., DeLalio, L. J., Johnstone, S. R. and Isakson, B. E. (2016) Small interfering RNA-mediated connexin gene knockdown in vascular endothelial and smooth muscle cells. In: Vinken, M. and Johnstone, S. R. (eds.) Gap Junction Protocols. Series: Methods in molecular biology, 1437. Springer New York, pp. 71-82. ISBN 9781493936625 (doi: 10.1007/978-1-4939-3664-9_5)

Abstract

Global knockout of vascular connexins can result in premature/neonatal death, severe developmental complications, or compensatory up-regulation of different connexin isoforms. Thus, specific connexin gene knockdown using RNAi-mediated technologies is a technique that allows investigators to efficiently monitor silencing effects of single or multiple connexin gene products. The present chapter describes the transient knockdown of connexins in vitro and ex vivo for cells of the blood vessel wall. In detail, different transfection methods for primary endothelial cells and ex vivo thoracodorsal arteries are described. Essential controls for validating transfection efficiency as well as targeted gene knockdown are explained. These protocols provide researchers with the ability to modify connexin gene expression levels in a multitude of experimental setups.

• Item Type: Book Sections
• Additional Information: This work was supported by National Institutes of Health grants (HL088554) and HL120840 and National Institutes of Health training grants (HL007284) (BEI) and by a University of Glasgow, Lord Kelvin Adam Smith Fellowship (SRJ).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Johnstone, Dr Scott
• Authors: Good, M. E., Begandt, D., DeLalio, L. J., Johnstone, S. R., and Isakson, B. E.
• College/School: College of Medical Veterinary and Life Sciences > School of Cardiovascular & Metabolic Health
• Journal Name: Methods in Molecular Biology
• Publisher: Springer New York
• ISSN: 1064-3745
• ISBN: 9781493936625