Ascarosides comprise a family of small signaling molecules that have been shown to regulate important events and behaviors in the life history of the nematode Caenorhabditis elegans. Although the different roles of individual ascarosides appear to be determined by the variances in chemical structure, the mechanisms by which ascarosides are synthesized as well as the locations in which ascarosides are produced within the worm are largely unknown. In this thesis, we examined ascaroside production in the intestine, hypodermis, and body wall muscle of the worm by driving the expression of the protein DAF-22 under different tissue-specific gene promoters. While the body wall muscle and hypodermis are capable of synthesizing ascarosides, the intestine appears to be the major site of pheromone production. Additionally, we found through transgenic rescue and HPLC-MS analysis, that the acyl-CoA synthetase ACS-7 plays a significant role in the addition of moieties derived from primary metabolic pathways to the 4’-position of the ascarylose sugar core of ascr#9.</p