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Understanding the Genetic Basis for piRNA Silencing in the Soma and Germline of Caenorhabditis elegansPeng, Yuli 07 1900 (has links)
C. elegans is a commonly used genetic model organism due to the ease of genetic
screens, transgenesis, and microscopy. Here, I describe methods that improve
transgenesis in C. elegans and the development of a genetic screen to identify genes
involved in the piRNA pathway. Transgenesis is commonly used for most laboratories
that utilize C. elegans and improvements are therefore likely to facilitate research across
many research areas. In the first chapter, I characterized a pan-muscular promoter that
drives fluorophore expression to help identify C. elegans transgenesis. This promoter is
an improved co-injection marker as it drives bright fluorescence with low toxicity and
high efficiency.
In the second chapter, I study piRNAs which are a large class of non-coding RNA that
play important roles in protecting the genome from transposable elements in most
animals. The study of piRNAs has mostly focused on their function in the germline, but
recent evidence suggests functions in somatic cells such as neurons. To identify genes
involved in the piRNA pathway in C. elegans, I performed a chemical genetic screen. I
identified one mutant with a somatic phenotype and six mutants with a germline
phenotype. I have focused on the germline and sequenced two strains and identified
candidate genes involved in the piRNA pathway. Future work will focus on validating
and identifying the remaining mutants.
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