Current Projects

     Our research workflow is diagramed in the figure below and remains our overarching strategy. From the most recent forward genetic screen, we identified 12 new alleles in 11 genes critical for neural patterning. We characterize each mutant phenotype in vivo and derive mouse embryonic fibroblasts (MEFs) so we can identify cellular phenotypes in vitro. We generate or obtain floxed null alleles of the genes we identify, so that we can understand the tissue and temporal roles of the gene. The floxed alleles also provide a powerful cell culture system in which we can delete the gene at will and add back variants to test the function of each. These variants are informed by published information describing the biochemical properties of each gene product as well as patient alleles. Fromour testing of variants in in vitro analyses (eg, ciliogenesis, Shh transcriptional responses), we subsequently select a few mutations for mouse model generation. This enables us to develop and refine hypotheses efficiently using the in vitro results and to study select variants in vivo for physiologically relevant data.

 

 

 

Figure 5: Pax3-GFP Expression in Mutants

Heritable lines displaying aberrant Pax3-GFP expression. These are the lines that display either abnormal (M2 and AB4) or a lack (AM3) of Pax3-GFP expression in the neural tube at embryonic day 10.5.