CHARACTERIZATION OF GENES ASSOCIATED WITH ABSENCE OF GERM LINE CELLS IN ZEBRAFISH (DANIO RERIO)
Carina Ramos, Kellee Siegfried.
University of Massachusetts Boston, Boston, MA.
Zebrafish is a crucial animal model to study early development and gene function in vertebrates. We are using zebrafish to identify and study genes that are necessary for germ line development. Our goal is to understand the role played by two genes in the germ cells: C17orf104 (named moto), and slc34a1a (named vacant) in zebrafish. By comparing the gene sequence of c17orf104 and slc34a1a from diverse species, we learned they are highly conserved in vertebrates, including Homo sapiens. Also, they are highly expressed in the reproductive system. Although, there is still a lack of information about the function of these 2 genes in germ cells. Our histological studies have shown mutations affecting these 2 genes in zebrafish, both ultimately resulting in sterile males. The mutation on C17orf104 is presumed to be responsible for a failure in meiosis. The mutation affecting slc34a1a leads to loss of all germ cells. In this study, we are testing how the genetic mutation at each locus affects the development of the germ line cells in zebrafish. To confirm that each of our original mutations is the cause of the observed sterility, we are generating new mutations in each. To this end, the CRISPR - associated proteins (Cas) system was used as a targeted genome mutagenesis tool. Furthermore, functional elements may be recognized on the basis of their evolutionary conservation.