Ribozyme- and siRNA-mediated suppression of RGS-containing RhoGEF proteins

Qin Wang, Min Liu, Tohru Kozasa, Jeffrey D. Rothestein, Paul C. Sternweis, Richard R. Neubig

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Given recent efforts to determine the sequence information on thousands of genes in the human genome, the current challenge is to identify the functions of these genes, including those encoding the regulator of G-protein signaling protein gene superfamily, and to establish their roles in particular signaling pathways in a native system. Increasingly, reverse genetic approaches are being used to address these questions. This article compares two powerful approaches [ribozyme and "short interfering" RNA (siRNA) techniques] under identical conditions for the first report on the suppression of endogenous RGS domain-containing RhoGEFs. The siRNA technique was found to be much more potent than ribozyme targeting at the same mRNA site of RGS-RhoGEFs. Also, the three siRNAs targeting LARG, PDZ-RhoGEF, and p115-RhoGEF are able to discriminate the closely related sequences within this RGS-RhoGEF gene family.

Original languageEnglish (US)
Pages (from-to)244-265
Number of pages22
JournalMethods in Enzymology
Volume389
DOIs
StatePublished - 2004

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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