FUNCTIONAL_ANALYSIS_OF_SF3B1_K700E_AND_SRSF2_P95H_MUTATIONS

The aim is that of investigating the consequences of SF3B1 K700E mutation in haematopoiesis by modeling it in the mouse animal model system. Because this mutation is thought to be gain-of-function and is always heterozygous, my approach is that of a heterozygous knock-in in mouse ES cells. SF3B1 is a ubiquitous protein with a crucial role in cell survival, so we predict that constitutive expression of the mutation in ES cell could be incompatible with cell survival/embryo development. For this reason, we want to create a conditional alleles that will only allow expression of the mutated proteins in adult life in the haematopoietic tissue. Human and mouse SF3B1 genes share 100% identity at the protein level, so I predict the functional consequences to be the same. Because SF3B1 encodes a splicing factor protein, I predict the functional consequences of its mutation could involve mis-splicing of RNA transcripts and lead to altered expression levels of key genes. I intend to use ES cells RNA to investigate what RNA species are mostly affected.