The overall aim of this project is to use an unbiased global approach to identify allfunctionally important in vivo binding sites for three major splicing regulatory proteins,SRSF1, SRSF5 and hnRNPA1 in human disease genes. We will investigate the synergisticand competitive effects, as well as the overlap in binding motifs of these proteins bydetermining global maps for each protein both in the presence of the other proteins and intheir absence. We will employ both the new and very powerful High-ThroughputSequencing of RNA isolated by Crosslinking ImmunoPrecipitation (HITS-CLIP) methodthat enables identification of the binding sites for the splicing regulatory protein in livingcells and also use next generation sequencing of RNA (RNA-seq). We will use cells wherewe can tightly control the expression of the individual splicing regulatory proteins. Bycombining inducible expression of the individual splicing regulatory proteins withsimultaneous knock down of the other splicing regulatory proteins, we will investigate theircomplex interplay and the dynamics of their binding to specific and shared regulatorymotifs.
|Effektiv start/slut dato||01/05/2012 → 01/01/2014|