Abstract
DNA vectors that express short hairpin RNAs (shRNAs) provide a new tool for reverse genetic analysis for selective long-term reduction of gene expression in mammalian cells. Using shRNA constructs with a cytomegalovirus promoter and an actin intron between the hairpins for stabilization, we reduce expression of an exogenously expressed gene, GFP and the endogenous protein, Translin-associated factor X (TRAX), in stably transfected Hela cell lines. The reduction of TRAX in Hela cells causes reduced cell proliferation. This decrease is specific as there is no equivalent reduction of the TRAX interacting protein, Testis brain RNA-binding protein, or any significant increase in a number of interferon-related target genes.
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