Study of the Endogenous CRAC Channel Using shRNA-Mediated Gene Silencing

Xuexin Zhang, Amy M. Spinelli, Wei Zhang, Mohamed Trebak

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The Ca 2+ release-activated Ca 2+ (CRAC) current is a major signaling event in non-excitable cells whereby Ca 2+ store depletion activates Ca 2+ entry across the plasma membrane from the extracellular space. Stromal interaction molecule 1 (STIM1) and Orai1 proteins are the key molecular players of the CRAC channel. Previous studies have linked activity of this channel to many physiological functions, and dysregulation of the CRAC channel has been associated with various diseases. In the absence of inducible tissue-specific knockout mice, in vivo knockdown studies examining the endogenous function of CRAC channel proteins, STIM1 and Orai1, are a challenge. In this chapter, we describe a lentiviral delivery system of shRNA-mediated gene silencing that has proven successful in studying the endogenous CRAC channel in vivo.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages137-145
Number of pages9
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1843
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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    Zhang, X., Spinelli, A. M., Zhang, W., & Trebak, M. (2018). Study of the Endogenous CRAC Channel Using shRNA-Mediated Gene Silencing. In Methods in Molecular Biology (pp. 137-145). (Methods in Molecular Biology; Vol. 1843). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8704-7_12