Research objectives and content
Despite a wealth of data coming from in vitro transcription and transfection experiments the role of TFIID subunits (TAFIIs in trancriptional regulation in mammalian cells has not been strictly defined. I propose to use mammalian genetic techniques to address this question. Previous studies have implicated TAFII28 as a cell-specific TAFlI involved in transcriptional regulation by nuclear receptors. I will determine the mRNA and protein expression pattern of TAFII28 in established cell lines and in the developing and adult mouse. According to the results of this study, I will use the technique of conditional cre/lox mediated homologous recombination to inactivate TAFII28 either in F9 embryonal carcinoma cells or in ES cells allowing the generation of TAFII28 null mice. The phenotype of the resulting TAFII28 null F9 cells and/or mice will be characterised. Training content (objective, benefit and expected impact)
The research I propose will enable me to learn important genetic techniques for the study of genes in mammalian cells and techniques in biochemistry. At the IGBMC I will gain important experience in the field of transcriptional regulation, embryology, and in the biology of nuclear receptors. The proposed research entails the use of homologous recombination in F9 cells, a procedure in which the IGBMC has considerable experience. I consider that the opportunity to learn these techniques at an outstanding institute such as the IGBMC is fundamental for my research carreer.
Links with industry / industrial relevance (22) None