Download Protein components of the haematopoietic gene regulatory network

 Protein components of the haematopoietic gene regulatory network RESEARCH PROJECT CODE -­‐ BIOL1691015 Haematopoietic Stem Cells (HSCs) have a long history in therapy but the current aim is their use in personalised medicine. To exploit them fully we must understand the processes that generate them and control their differentiation using a GRN, which can be used to predict the behaviour of cells in response to treatment. The key molecules driving the generation and differentiation of HSCs are identified but the detailed mechanisms controlling them remain unclear. This is for two reasons: the limited ability to identify these cells unequivocally in live embryos and the lack of antibodies recognising key haematopoietic TFs in the only model organism where the key HSCs are accessible, Xenopus. We have prepared these antibodies and are characterising Xenopus lines in which the HSCs are labelled fluorescently. The student will first develop the GRN for the ventral blood islands (VBI), which can be dissected from the embryo using anatomical landmarks and already much of the data to generate the GRN are in place; however gain and loss of function experiments (GOF/LOF), in situ hybridisation analysis and western blotting are still required to fill the gaps. ChIP will be used to test whether the interaction between TF and target gene initially revealed in the GOF/LOF analysis is direct; this is essential to improve the accuracy of the GRN. Using this experience the student will then analyse the adult HSCs in the dorso-­‐lateral plate mesoderm (DLP). Here the GOF/LOF experiments are almost complete, but nothing is known about the expression of key proteins in these cells nor have any of the TF targets been confirmed by ChIP; the student will add these data to the haematopoietic GRN in the DLP. Training will be in embryology (including GOF/LOF), antibody staining, in situ hybridisation, bioinformatics and ChIP. Informal enquiries can be made to Prof Matt Guille on 02392 842047 or [email protected]. Key words: stem cells; Xenopus; haematopoiesis; genetic regulatory networks; genome editing When applying please note the project code: BIOL1691015