Submission Title

Exploration of the Role of the CCR4-NOT Deadenylase Complex Subunit Regena/NOT2 in MicroRNA-Mediated Gene Silencing

Location

Jereld R. Nicholson Library: Grand Avenue

Subject Area

Biology

Description

Our goal is to determine the cellular mechanisms that regulate microRNA (miRNA)-mediated gene silencing. MiRNAs are non-coding RNA molecules that interact with target-gene messenger RNA transcripts via complementary base pairing and silence target gene expression via interaction with the miRNA-induced silencing complex (miRISC). A forward genetic screen was carried out to identify requirements for miRNA-mediated silencing in Drosophila melanogaster. miRNA function was assayed using Green Fluorescent Protein (GFP)-based reporters of gene silencing. The GFP transgenes contain miRNA binding sites and are regulated by endogenous miRNAs. We have genetic evidence demonstrating a role for Regena (NOT2), a subunit of the CCR4-NOT deadenylase complex, in miRNA-mediated gene silencing. We are investigating the necessity of this gene in miRNA-mediated gene silencing at various stages of Drosophila development by determining differences in gene expression in mutant larva lacking functional NOT2. We have shown NOT2-mediated reporter silencing to indeed be dependent upon the activity of miRNAs, as demonstrated by the use of an additional GFP-based reporter of gene silencing. Reporter gene expression in larval fat body tissues is being assessed to determine whether NOT2 is required for silencing at different developmental time points and in different tissues of Drosophila.

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Exploration of the Role of the CCR4-NOT Deadenylase Complex Subunit Regena/NOT2 in MicroRNA-Mediated Gene Silencing

Jereld R. Nicholson Library: Grand Avenue

Our goal is to determine the cellular mechanisms that regulate microRNA (miRNA)-mediated gene silencing. MiRNAs are non-coding RNA molecules that interact with target-gene messenger RNA transcripts via complementary base pairing and silence target gene expression via interaction with the miRNA-induced silencing complex (miRISC). A forward genetic screen was carried out to identify requirements for miRNA-mediated silencing in Drosophila melanogaster. miRNA function was assayed using Green Fluorescent Protein (GFP)-based reporters of gene silencing. The GFP transgenes contain miRNA binding sites and are regulated by endogenous miRNAs. We have genetic evidence demonstrating a role for Regena (NOT2), a subunit of the CCR4-NOT deadenylase complex, in miRNA-mediated gene silencing. We are investigating the necessity of this gene in miRNA-mediated gene silencing at various stages of Drosophila development by determining differences in gene expression in mutant larva lacking functional NOT2. We have shown NOT2-mediated reporter silencing to indeed be dependent upon the activity of miRNAs, as demonstrated by the use of an additional GFP-based reporter of gene silencing. Reporter gene expression in larval fat body tissues is being assessed to determine whether NOT2 is required for silencing at different developmental time points and in different tissues of Drosophila.