Transgene Expression Localized During Stages of Spermatogenic Development

Sperm malformed during spermatogenesis may cause infertility and lead to developmental mutations. Essential for viable sperm formation, GSP-3 and GSP-4 genes encode for Glc7/PP1 phosphatase–they are uniquely expressed during spermatogenesis, and function during sperm meiosis and sperm motility generation. It is unknown how GSP-3 and GSP-4 change localization to function in these different processes. Expression of fluorescently-tagged transgenes localized in the gametes creates a novel opportunity for investigating GSP-3 and GSP-4 functions in sperm formation. Transgene expression in C. elegans can be achieved by employing the Mos1 mediated Single Copy gene Insertion (MosSCI) injection technique. We will construct a MosSCI DNA vector containing a GSP-3 promoter, the GSP-3 open reading frame (ORF), and the 3' untranslated region (3'-UTR) fused to a fluorescent GFP protein construct. This will be injected into C. elegans to produce progeny that express labeled transgenes that can be viewed in a living animal under a fluorescent scope. We expect that this tool can be employed for future investigation of spermatogenesis.