Silencing the cardiac potassium channel Kv4.3 by RNA interference in a CHO expression system

Cotella, Diego and Jost, Norbert and Darna, Mahesh and Radicke, Susanne and Ravens, Ursula and Wettwer, Erich (2005) Silencing the cardiac potassium channel Kv4.3 by RNA interference in a CHO expression system. Biochemical and Biophysical Research Communications, 330 (2). pp. 555-560. ISSN 0006-291X

Text 1109505.pdf Restricted to Registered users only Download (554kB) | Request a copy

Abstract

RNA interference (RNAi) is a powerful technique for gene silencing, in which the downregulation of mRNA is triggered by short RNAs complementary to a target mRNA sequence, with consequent reduction of the encoded protein. The aim of this study was to test the effects of silencing the expression of the cardiac potassium channel Kv4.3 in a heterologous expression system, in order to investigate the effect of RNAi on channel properties. A Chinese hamster ovary cell line stably expressing Kv4.3 and the accessory β-subunit KChIP2 was transfected with small-interfering RNAs (siRNAs) targeting Kv4.3. Effects of RNAi were monitored at the mRNA, protein, and functional levels. Real-time PCR and immunofluorescence staining revealed significant reduction of Kv4.3 mRNA and protein expression. These results were confirmed by functional patch-clamp measurements of the transient outward current (Ito) which was reduced up to 80% by RNAi. We conclude that the use of siRNAs reagents for post-transcriptional gene silencing is a new effective method for the reduction of the expression and function of different ionic channels which may be adapted for studying their role also in native cells.

Actions (login required)

Edit Item