THE WORM, THE MYTH, THE LEGEND QUANTIFICATION OF MALE rha-1 MUTANT GERMLINE GENE EXPRESSION REGULATED BY SMALL RNA PATHWAYS OF Caenorhabditis elegans

Date of Award

2013

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Walstrom, Katherine

Keywords

C. Elegans, RNA Interference, qRT-PCR

Area of Concentration

Biochemistry

Abstract

RNA Helicase A (RHA-1) is a conserved protein thought to be involved in the regulation of germline gene expression in C. elegans. Previous studies involving mutants lacking RHA-1 exhibited a temperature sensitive sterility defect similar to eri-1 mutants. ERI-1 is an extensively studied protein known to have a key role in germline gene regulation through RNA interference, wherein small segments of RNA suppress gene expression. Since ERI-1 has a regulatory role in 26G RNAi, and eri-1 mutants exhibit a similar phenotype as rha-1 mutants, it was postulated that RHA-1 may also have a role in the eri-1-dependent RNAi pathway. RHA-1 appears to affect spermatogenesis in particular, so male him-8 mutants were used to allow for an examination of worms that produce sperm for a longer period of time. Through the use of quantitative real-time RT-PCR (qRT-PCR), several genes thought to be regulated by the 26G and 21U RNAi pathways were tested in mutant strains lacking RHA-1, ERI-1, or both RHA-1 and ERI-1. It was found that rha-1 mutants do not have significantly altered gene expression of the genes tested that are regulated by the 26G pathway. Although RHA-1 does not appear to significantly affect gene expression levels tested, there is still evidence that it plays a regulatory role in spermatogenesis-related processes, given previous phenotype observations. There is still a possibility that it may be involved in alternate RNAi pathways.

Rights

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