Training objectives and contentMyft-5 is the first myogenic regulatory factor (MRF) expressed during mouse embryogenesis and plays a central role in skeletal muscle determination. Myf-5 is initially expressed in the most dorsal and medial cells of the dermamyotome, the location of the epaxial muscle precursors. Transgenesis experiments have shown that myf-5 has a separate enhancer for each of the anatomically distinct muscle precursor regions in the embryo. Deletion of a 2.6kb segment close to the adjacent MRF4 gene eliminates expression of myf-5 in the epaxial domain, while not altering any other aspect of the pattern.
We project to: -Investigate whether this 2.6kbsegment is sufficient for epaxial expression and define the minimal DNA sequences required. -Define protein-binding sites in the minimal region and test their function by mutagenesis and transgenesis. Characterise the proteins required for enhancer function. -Test putative signalling molecules involved in myogenesis by crossing epaxial enhancer-reporter constructs with the appropriate knockouts. -Use constructs containing both myf-5 and MRF4 genes, and carrying two reporter genes to investigate whether there are sequences that regulate both MRF genes. Training content (objective, benefit and expected impact). My expertise is in Cellular Biology and in vitro techniques. This project involves an extensive training in transgenesis, mouse breeding, micro-dissection and Developmental Biology. Links with industry / industrial relevance (22)