Building a neurogenic gene regulatory network for C. teleta will facilitate comparisons with other bilaterian groups and help elucidate how central nervous systems evolved. To continue identifying genes involved in early neurogenesis in C. teleta, spatial and temporal expression patterns for candidate neurogenic genes will be generated using in situ hybridization and quantitative PCR. Candidates will then be tested using functional knock-down approaches (morpholino and siRNA injection).
To identify additional putative downstream targets of CapI-Ash1, microarray or 454 sequencing analysis will be used to compare normal versus depletion (CapI-ash1 morpholino) or ectopic expression (CapI-ash1 mRNA) conditions. Putative targets will be verified by quantitative PCR and in situ hybridization after morpholino or mRNA injection.