The family of NimA-related kinases (NEKs) are highly conserved in eukaryotes where they have been reported to regulate a number of diverse processes including mitosis, cytokinesis, DNA repair, ciliogenesis, and inflammation. Correspondingly, misregulation of NEKs have been linked to a variety of diseases including cancer and renal and cardiac disorders. Nevertheless, the targets of NEKs, as well as the components of NEK signaling pathways, remain largely unknown. In contrast to humans, which encode 11 NEK family members, the C. elegans genome encodes 4 family members (NEKL-1/NEK9, NEKL-2/NEK8, NEKL-3/NEK6/7, and NEKL-4/NEK10). We recently reported a novel function for NEKL-2 and NEKL-3 in regulating the completion of C. elegans molting during larval development. Our published and recent studies indicate that incomplete molting stems, at least in part, from defects in endocytosis in the apical epidermis of C. elegans larvae, thus implicating NEKs in process associated with intracellular trafficking. To gain further insight into the molecular, cellular, and developmental functions of NEK kinases, we have undertaken a number of approaches including genetic analysis. Using CRIPSR/Cas9 methods, we have used an iterative approach to generate a range of loss-of-function (LOF) alleles of nekl kinases We find that weak hypomorphic alleles in nekl-2 and nekl-3, which display no defects as single mutants, are synthetically lethal, leading to near uniform larval arrest in nekl-2; nekl-3 double mutants. We reasoned that nekl-2; nekl-3 may provide an optimal background for identifying suppressors of NEKL-associated molting defects given that the individual nekl alleles are weak LOFs and that suppression of lethality could be incurred by mutations that affect either the NEKL-2 or NEKL-3 pathways or both. Using a semi-clonal F1 screen, we were successful in identifying ~12 suppressor mutations of varying strengths and we will describe progress on their molecular identification. We will also describe a non-clonal screen, that makes use of a counter-selectable marker in C. elegans, to identify additional nekl-2; nekl-3 suppressors. This method could be adapted to any suppressor screen where the lethal phenotype of the parental starting strain is incompletely penetrant. Finally, we will discuss focused RNAi approaches to directly identify protein kinases and protein phosphatases that modulate signaling by NEKL kinases.