Abstract
Sleep has been observed in all animals with a nervous system, however, the neural and molecular mechanisms that regulate it are not fully understood. The genetically tractable nematode Caenorhabditis elegans has a simple and fully mapped nervous system and has emerged as a powerful sleep model. C. eleganssleeps concurrently with each larval molt, called developmentally timed sleep (DTS), which is timed by the PERIOD(PER) homolog LIN‐42. C. elegans also sleep during adulthood in response to cellular damage incurred by stressful stimuli, called stress‐induced sleep (SIS). Based on LIN‐42’s central role in the timing of DTS, we sought to examine its role in the timing of SIS. Null mutations of lin‐42are lethal, so to study the requirement of LIN‐42 during SIS, we used CRISPR/Cas9 to insert the auxin‐induced degron and green fluorescent protein sequence in frame with each isoform of the lin‐42 gene. In the absence of auxin these worms are viable and grow successfully to adulthood. We are currently constructing transgenic strains that will allow us to degrade the LIN‐42 in different subsets of cells. To do this we are expressing the coding sequence for the auxin perceptive F‐box protein TIR1 from different tissue‐specific promoters, including those expressed in all cells, and specific sleep‐regulating neurons. Once these strains are constructed we will degrade LIN‐42 protein and measure the effects on SIS.
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