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

Diego Cotella, Norbert Jost, Mahesh Darna, Susanne Radicke, Ursula Ravens, Erich Wettwer

Research output: Contribution to journalArticle

6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)555-560
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume330
Issue number2
DOIs
Publication statusPublished - May 6 2005

Keywords

  • CHO cell
  • Gene silencing
  • I
  • Ion channels
  • Kv4.3
  • RNA interference
  • siRNA

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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