A fluorescent reporter detecting precise homologous recombination transgenesis activity

The broad long term objective and specific aims of the fast track SBIR research are to create a reagent RecombiRED for use in microinjection of embryos for identifying a subset of embryos as recombination competent injections The ability to quickly identify genome editing events in an embryo allows researcher to quickly make precision animal models of human disease states Precise animal models allow for greater scientific understanding of a human disease and provides a robust platform for drug discovery screening The RecombiRED reagent relies on induced fluorescence in response to activation of homologous recombination repair within the embryo Detection of homologous recombination repair in injected embryos with RecombiRED allows animal husbandry and further screening efforts to be restricted to a subset of embryos by up to x for having a desired precise knockin of new genetic material at a specific genetic locus The phase I research adapts a the RecombiRED reagent developed in C elegans model organism to have the appropriate expression control elements enabling sufficient activity in zebrafish embryo In Phase II the additional research further optimizes parameters of the RecombiRED reagent to minimize false positive and false negative readouts which then allows maximal forecasting of targeted genome edit events Finally a set of real world transgenesis projects are used as premarket beta test to demonstrate the ubiquity and robustness of the RecombiRED for simplifying discovery of genome editing in the creation of animals models for use in drug discovery The relevance of this research to public health is the creation of an experimental tool for simplifying the discovery of genetically altered model organisms The tool allows faster creation of animal models for use in better scientific discovery and faster drug development