Location

Jereld R. Nicholson Library

Subject Area

Biology

Description

Mature microRNAs (miRNA) are ~22 nucleotide long single-stranded ribonucleic acids essential for gene silencing. Silencing occurs when miRNAs are processed via endonucleolytic cleavage and subsequently associate with the miRNA-induced silencing complex (miRISC). miRISC binds via complementary base pairing to target mRNAs, and target mRNAs are silenced by either mRNA degradation, translational block, or both. Knowledge of all genes required for silencing is incomplete.

We aim to determine the molecular mechanism of silencing by identifying and characterizing genes required for silencing. A forward genetic screen was performed using EMS mutagenesis of Drosophila melanogaster to generate mutant lines with disrupted gene silencing as visualized by a GFP-based fluorescent reporter of silencing. Locations of EMS-induced mutations are being mapped by determination of recombination frequencies between these mutations and molecularly defined P-element insertions. Preliminary recombination mapping reveals that our mutation of interest (I1-5) is found within a discrete region of the genome on chromosome 3R. A new fly line has also been generated to assist with this preliminary recombination mapping. Future deficiency mapping and complementation tests combining the mutation and alleles of candidate genes will reveal the location of our mutation, and lead us to identify a gene required for micro-RNA mediated gene silencing.

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May 15th, 9:30 AM May 15th, 10:45 AM

Identifying a New Gene Required for microRNA-mediated Gene Silencing in Drosophila melanogaster

Jereld R. Nicholson Library

Mature microRNAs (miRNA) are ~22 nucleotide long single-stranded ribonucleic acids essential for gene silencing. Silencing occurs when miRNAs are processed via endonucleolytic cleavage and subsequently associate with the miRNA-induced silencing complex (miRISC). miRISC binds via complementary base pairing to target mRNAs, and target mRNAs are silenced by either mRNA degradation, translational block, or both. Knowledge of all genes required for silencing is incomplete.

We aim to determine the molecular mechanism of silencing by identifying and characterizing genes required for silencing. A forward genetic screen was performed using EMS mutagenesis of Drosophila melanogaster to generate mutant lines with disrupted gene silencing as visualized by a GFP-based fluorescent reporter of silencing. Locations of EMS-induced mutations are being mapped by determination of recombination frequencies between these mutations and molecularly defined P-element insertions. Preliminary recombination mapping reveals that our mutation of interest (I1-5) is found within a discrete region of the genome on chromosome 3R. A new fly line has also been generated to assist with this preliminary recombination mapping. Future deficiency mapping and complementation tests combining the mutation and alleles of candidate genes will reveal the location of our mutation, and lead us to identify a gene required for micro-RNA mediated gene silencing.

 

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