RNAi analysis of the FBP transactivator family

The FBP (FUSE Binding Protein) transcription factor family is composed of three highly related members; FBP/FBP1, FBP2, and FBP3. While they all share distinct modular protein structures and nucleic acid-binding specificities, their transactivator strength is graded and they also differ in their interaction with their protein partners. Especially from the extensive nuclear co-localization of these molecules, we anticipated that they co-regulate a set of common target genes. Thus we knocked down the level of each FBP using specific siRNA and obtained an initial assessment of the degree to which these factors act concertedly or independently. Keywords: binding site identification design Each FBP family member was knocked down by transiently transfecting siRNAs, and the total RNA was compared after two days against the control RNA from the negative control transfection. Six arrays were hybridized and analyzed for human primary skin fibroblast samples; two independent experimental sets, each for the three different FBPs.