We are developing an experimental system to measure cooperation between genes.  In order to interrogate a full range of dosage levels, a library of constitutive promoters has been produced through mutagenesis of the cytomegalovirus immediate early and elongation factor 1 alpha promoters.  The system can be used to drive a range of expression of several exogenous genes in a single, heterogeneous culture.  By co-expressing the exogenous genes of interest with fluorescent protein reporter genes, large numbers of cells in the heterogeneous populations can be quickly analyzed by flow cytometry.  In order to appropriately evaluate genetic cooperation, the expression library is coupled with a fluorescent phenotypic readout consisting of either genetic reporters or chemical assay.  In particular we are developing a fluorescent proliferation reporter containing cell cycle-responsive promoters allowing for correlation between fluorescent protein levels and proliferation rate.Ultimately, we aim to evaluate the effect of expression of various combinations and dosages of oncogenes and tumor suppressors on cellular proliferation.  The response from expression of individual genes can be used to determine additive response, the sum of individually expressed gene contributions.  Cooperative or antagonistic behavior is then determined by comparison of genes expressed in combination to the additive response.  This approach will allow not only for determination of important gene combinations in causing increased cellular proliferation, but also the optimal dosage ranges for such behavior.

System to evaluate genetic interactions using a constitutive promoter library

239th National Meeting (2010)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.