RNA interference and ion channel physiology

Authors: Palmer, Melissa¹; Fahrenkrug, Scott²; O'Grady, Scott³

Source: Cell Biochemistry and Biophysics, Volume 46, Number 2, June 2006 , pp. 175-191(17)

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Abstract:

RNA interference (RNAi), through expression of small, double-stranded RNAs or short hairpin RNAs, produces sequence-specific mRNA degradation and decreased gene expression. Since its discovery in 1998 (Fire et al., 1998, Nature 391, 806-811), RNAi has rapidly become one of the most widely used technologies for exploring gene function in eukaryotic cells. Although the topic of RNAi has been the subject of a large number of excellent reviews, the focus of this article is on its application to the study of ion channel physiology in animal cells. In this regard, RNAi has provided definitive identification of ion channel subtypes responsible for both basal and stimulated ion conduction across the plasma membrane of several cell types. The approach has been particularly effective in identifying and establishing the contribution of auxiliary subunits and regulatory proteins to the overall function of ion channel complexes. Moreover, selective knockdown of ion channel expression has been a valuable means of demonstrating roles in the development of specific cell domains and in the normal growth of certain cell types. In this review, a brief description of the general mechanism of RNAi is presented, followed by a discussion of some important considerations for the in vitro application of this technology and in producing transgenic animals as models for human disease. We then describe several examples of where RNAi has been used to investigate the physiological role of ion channels in cells from model organisms (Caenorhabditis elegans and Drosophila melanogaster) and in mammalian cells.

Keywords: RNAi; shRNA; siRNA; RNA silencing; gene silencing; ion transport

Document Type: Research article

DOI: http://dx.doi.org/10.1385/CBB:46:2:175

Affiliations: 1: Department of Physiology, University of Minnesota, 495 Animal Science/Veterinary Medicine Building, 1988 Fitch Avenue, 55108, St. Paul, MN, 2: Department of Animal Science, University of Minnesota, 495 Animal Science/Veterinary Medicine Building, 1988 Fitch Avenue, 55108, St. Paul, MN, 3: Department of Physiology, University of Minnesota, 495 Animal Science/Veterinary Medicine Building, 1988 Fitch Avenue, 55108, St. Paul, MN, Email: ograd001@umn.edu

Publication date: 2006-06-01

RNA interference and ion channel physiology

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