Deciphering the Molecular Mechanisms of Gene Silencing through the Genetic Analysis of Drosophila melanogaster
Biology | Genetics | Molecular Genetics
In an effort to elucidate the molecular mechanisms of gene silencing in Drosophila melanogaster, we have identified a role for the CCR4/NOT complex subunit unit Regena/NOT2 in microRNA-mediated gene silencing. A fluorescent sensor of microRNA-mediated gene silencing was used to evaluate the effect of a mutation in Regena/NOT2 on silencing during early stages of Drosophila development. This evaluation was made possible by the development of reagents and protocols for evaluating gene silencing in Drosophila larva. Further, a combination of recombination and deficiency mapping was followed by the amplification and sequencing of the Regena/NOT2 gene from our mutant lines. Sequence analysis has revealed a nonsense mutation in the Regena/NOT2 gene. The reagents and protocols developed in this work have also been used to evaluate the role of perturbations in the insulin signaling pathway on gene silencing during early stages of Drosophila development. In addition, preliminary recombination mapping has been carried out to identify another novel gene required for microRNA-mediated gene silencing. Taken together, this work identifies new genes not previously demonstrated to be essential for gene silencing, and helps to further describe the molecular mechanisms of microRNA-mediated gene silencing.
Reinke, Catherine, "Deciphering the Molecular Mechanisms of Gene Silencing through the Genetic Analysis of Drosophila melanogaster" (2014). Post-Grant Reports. Report. Submission 50.