Phenotypic Analysis Of Human Disease Gene RRAGB Mutant Allele In Drosophila Melanogaster

Phenotypic analysis of human disease gene RRAGB mutant allele in Drosophila melanogaster. Abstract. Introduction. MTORC1- Rag GTPase. Generation of a knockout mutant by using the CRISPR/Cas9 system in a Drosophila melanogaster. Reference.

Drosophila melanogaster Ras related GTP binding A/B gene (RagA-B) is orthologous to the human disease gene RRAGB. RRAGB encodes Ras related GTP binding protein B that is essential mediators of amino acid signals to mTORC1. It relays amino acid signals by recruiting mTORC1 signaling cascade to the lysosome and regulates cell growth, size and viability. mTORC1 is a highly conserved essential complex composed of mTOR (mammalian target of rapamycin) and its regulators in all eukaryotes. Due to the crucial role of mTORC1 in the cell regulation, RRAGB SNP mutation (T271S) seems to cause a human rare undiagnosed disease. To understand how the pathophysiology of RRAGB mutation in human, I set out to perform phenotypic analysis of the RRAGB mutation in Drosophila melanogaster as a model organism. I generate a null mutant allele of RagA-B using CRISPR/Cas9 mediated mutagenesis. After generating the stable mutation line by crossing with balancer and Gal4-UAS system using BAC recombineering in Drosophila, I rescue the mutant with transgenes that carry mutant or wild type form of the human homolog RRAGB. With these mutant and rescue strains, I will be able to analyze the detailed phenotypic effect of the human disease mutant of RRAGB on cell number and size of wing in Drosophila melanogaster. These experiments can suggest some biological information of RRAGB which is conserved disease gene both in human and flies. Although it is not perfect approach to the human disease, it can make positive feedback on human medical genomics and bioinformatics.