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Show THE UNIVERSITY OF UTAH RESEARCH POSTERS ON THE HILL 2013 INTEGRATING THE FLP RECOMBINAS SYSTEM INTO THE C.ELEGAN GERMLINE Rosalie Griffin (M.Wayne Davis, ErikJorgensen) Department of Biology University of Utah H Integrating the FLP Recombinase System into the C. elegans Germline Rosalie Griffin, Wayne Davis, Erik Jorgensen Department of Biology u C elegans is an excellent model organism. THF. UNIVERSITY OF UTAH limited use of FLP in C. elegans has been successful. HHMI Results and Future Direction Greater genomic control is desired in C elegans. Site-specific recombination provides greater control. « and L n o d o i t ( S k a m n 2011) >tt allow ui to study rhe cflca « To provide trans-generational change, w e will express the FLP recombinase system in the germline of C. elegans. .*_ . •~vrr^*- tmr S v O v n . Rippas* (FLP) cataryzn e n specific target sequence* m O N A c Caenorhabditis elegans is an excellent genetic model organism for the study of the complete nervous system, including genes involved in development, neuronal function and behavior. By observing the effect of removing a gene, w e can infer the function of the protein that the gene encodes. However, current methods, which create random null mutations, have limited control over the study of gene function. Greater control over the genome has been achieved in Drosophila and mouse through site-specific recombination such as FLP recombinase. Flippase (FLP) catalyzes recombination between specific target sequences in D N A called FLP recognition targets (FRT). FLP binds to FRT regions and recombines out any D N A between the target sequences. Previously, the FLP recombinase system has been successfully used in C. elegans to spatially and temporally control gene activity. Such systems, however, only affect the somatic cells of the animal. To provide trans-generational change w e propose to express the FLP recombinase system in the germline of C. elegans. W e have generated a plasmid that contains flippase under the control of a germline promoter in a Mosl transposon vector. W e have generated a second plasmid that contains FRT sites flanking a transcription terminator inside a wild-type copy of the unc-119 gene, unc-119 mutant animals have an easy to recognize uncoordinated phenotype.We have inserted the flippase plasmid and will insert the FRT plasmid into the genome of an unc-119 mutant animal. Eventual recombinant excision of the transcription terminator by flippase will allow the inserted wild-type copy of unc-119 to be transcribed, causing rescue to a wild-type phenotype in the worm. This phenotype rescue will indicate the FLP recombinase system is working in the germline. Successful integration of the FLP recombinase system in the germline of C. elegans will provide a powerful new tool for the study of genes. 15 |