Interaction of the RNA binding protein Spen with Xist RNA (360G-Wellcome-202461_Z_16_Z)

My research will look into the role of the transcriptional repressor Spen in X inativation. Spen has already been found to be associated with the long non-coding RNA Xist that is known to mediate X inactivation by coating the surface of the inactive X chromosome. The nature of the interaction of Spen with Xist has not been investiagted in detail - the goal of my project is to characterise this interaction. My project will examine the importance of an RNA recognition motif (RRM) domain on the Spen protein, investigating its role in Xist binding. I will generate a mutant construct of Spen with the RRM domain of interest deleted, and examine its colocalisation with Xist. Xist will be expressed from an inducible promoter. The results of my project will generate data that will help to ascertain the importance of the RRM of Spen for Xist binding, helping to provide insights into the mechanism of X inactivation.