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. elegans sleeps 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-42 are 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.
Support or Funding Information
National Science Foundation Grants IOS-CAREER-1845020 and DBI-MRI-1919847
