Since its discovery, RNA interference (RNAi) has become a powerful tool used in laboratories to understand developmental processes, and it is now commonly used to investigate gene function. Using RNAi, scientists can routinely knock out specific genes to prevent their expression, observe its phenotypes, and then help determine the function of a gene. RNAi offers enormous potential, not only as a tool in biological research but also as a therapeutic approach for silencing disease-causing genes. However, to achieve full potential for such a powerful tool, one must understand the mechanism behind it. In the last decade, great progress has been made in understanding the mechanisms of RNAi. Nevertheless, there are many questions that still remain to be answered. What factors are involved in RNAi? What are their roles in RNAi? To answer these questions, I have screened for genes that are involved in RNAi in Drosophila melanogaster. With the use of both genetics and biochemistry, I have identified some of the genes and characterized their functions. Using proteomics, I have identified list of genes that are possibly regulated by miRNAs. In the RNAi pathway, siRNAs and miRNAs direct Argonaute proteins within the RNA-induced silencing complex (RISC) to silence complementary or partially complementary mRNA targets by mRNA cleavage or translational repression, respectively. I have isolated an allele of Ago2 that specifically affects siRISC assembly showing in vivo and in vitro that Ago2 is required for efficient release of passenger strand siRNA. I have also demonstrated that mutant alleles of a novel gene, dFbxo11, and loqs affect both siRNA and miRNA pathways. Altogether, I have answered several key questions that were addressed to help explain the mechanisms of RNAi in Drosophila melanogaster.

Last modified

• 09/11/2018

Creator

• Kevin Kim

DOI

• https://doi.org/10.21985/N2P44M

Subject

• Interdepartmental Biological Sciences Program

Keyword

• General Biology

Date created

• 2008-05-12

Resource type

• Dissertation

Rights statement

• In Copyright