The interplay between transposons and piRNA pathways

项目摘要/摘要 转座子是一种大量的遗传寄生虫，渗透到人类基因组的45%，是主要的比例 在所有动物基因组中。我们的实验室研究了Piwi/piRNA途径，即RNA干扰(RNAi)的手臂 系统，识别和沉默转座子转录，以保持基因组的稳定性和繁殖力。我们的实验室 专门研究转座子和piwi/piRNA途径之间的相互作用和动力学 更好地学习如何防止转座子扩散并对动物健康造成负面影响的目标。 为了从根本上揭示动物基因组和转座子之间的调控机制，我们正在 在Piwi/piRNA途径上部署基因组学、生化和小RNA分析方法。 本项目的主要重点是研究一种高流动性的Har-P元件转座子变异体 发现了P-转座酶驱动性腺发育不全(GD)不孕综合征的弹药 果蝇。我们项目的优势依赖于独特的果蝇品系和细胞系分析 刘实验室研究Har-P元件如何受piRNAs调控，Flamenco piRNA的增强子如何 簇可以从Har-P元件产生piRNAs，Har-P piRNAs是如何调节内含子剪接的。 本项目将实现以下目标：目标1：确定动员能力和 Har-P变异体的父系沉默机制，这是一种导致严重GD的超移动转座子。 目的2：分析来自Flamenco piRNA簇的P元件piRNA的转录调控 以及piRNA沉默和内含子剪接抑制之间的机制联系。这个项目将产生新的 深入了解转座子调控，为研究生育综合征创造新的研究试剂和工具 并告知由piRNAs介导的跨代表观遗传过程。

Project Summary/Abstract Transposons are prolific genetic parasites infiltrating >45% of the human genome and are major proportions of all animal genomes. Our lab studies how the Piwi/piRNA pathway, an arm of the RNA interference (RNAi) system, recognizes and silences transposon transcripts to preserve genome stability and fertility. Our lab specializes in examining the interplay and dynamics between transposons and the Piwi/piRNA pathway with the goal of learning better how to prevent transposons from proliferating and negatively affecting animal health. To fundamentally uncover the regulatory mechanisms between animal genomes and transposons, we are deploying genomics, biochemical and small RNA analytical approaches on the Piwi/piRNA pathway. The major focus of this project is to study a hyper-mobile Har-P-element transposon variant that we discovered is the ammunition for P-transposase to drive the infertility syndrome of gonadal dysgenesis (GD) in Drosophila. Our project’s advantage relies on unique Drosophila strains and cell line assays developed in the Lau lab to study how the Har-P-element is regulated by piRNAs, how the enhancer of the Flamenco piRNA cluster can generate piRNAs from Har-P-elements, and how do Har-P piRNAs regulate intron splicing. This project will achieve the following aims in this project: Aim 1: Determine the mobilization capacity and paternal silencing mechanism of the Har-P-element variant, a hypermobile transposon that drives severe GD. Aim 2: Dissect the transcriptional regulation of P-element piRNAs coming from the Flamenco piRNA cluster and the mechanistic link between piRNA silencing and intron splicing inhibition. This project will yield new insight into transposon regulation, generate new research reagents and tools for studying the fertility syndrome of GD, and inform on transgenerational epigenetic processes mediated by piRNAs.