Tagging Juvenile Hormone receptors in Drosophila

Juvenile Hormone (JH) is a key component of the insect endocrine system that regulates many functions. In Drosophila, JH elicits its action through the paralogous bHLH receptors, Methoprene tolerant (Met) and Germ cell-expressed (Gce). An important aspect of receptor functioning lies in its subcellular localization, and it has been identified that receptor localization is regulated by both hormone dependent and independent mechanisms. To better understand localization, we generated two epitope tagged receptor fly lines using CRISPR/Cas9 technology. The receptors were tagged at their carboxy termini with different epitopes, Met with V5 and Gce with 3xFLAG. The generated models were confirmed for the expression of epitope tagged receptors through western blot, followed by immunostaining for studying the subcellular localization in various tissues at different developmental stages. We observed localization to be predominantly nuclear. These models will be used to study localization of receptors in a JH-deficient background to study hormone dependent localization of the receptors.