Understanding the architecture, regulation, and function of piRNA-producing genes

项目III.了解产生piRNA的基因的结构、调节和功能 菲利普·D·扎莫尔，私家侦探 项目摘要 在果蝇、哺乳动物和蠕虫中，PIWI相互作用的RNA(PiRNAs)使转座子和重复元件沉默， 保护生殖系基因组免受DNA损伤和突变，并确保遗传信息传递 世世代代忠贞不渝。它们还有其他鲜为人知的功能。我们建议 确定基因结构、基因组结构和小鼠、苍蝇、 还有虫子。然后，我们将使用这些信息来调查为什么一些长RNA制造piRNA，而另一些长RNA制造piRNA 不要这样做。我们还将探索全长的piRNA前体转录本，通常是数万个碱基长的 转化成只有23-35个核苷酸长的成熟的pIRNA。最后，我们将测试各种假设，这些假设由 我们的数据，关于piRNA如何发挥作用，特别是在哺乳动物中，大多数piRNA不对应于 转座子。为了实现这些目标，我们将结合遗传学、高通量测序和经典 生物化学。我们认为，通过研究三个关于piRNA途径的常见问题 动物模型，我们发现的保守和发散特征将使我们对此有更深的理解 这是一条古老的路径，而不是单独研究单一物种所能实现的。 相关性 被称为“piRNAs”的小RNA保护动物精子和卵子前体细胞免受自私基因的破坏。 被称为转座子或“跳跃基因”的元素。没有piRNA，从蠕虫到哺乳动物的动物都是不育的， 然而，我们不知道piRNAs是如何合成的，对许多piRNAs来说，它们调控的是什么。我们希望能获得 对这一令人惊叹的过程有了更完整、更深刻的理解。

Project III. Understanding the architecture, regulation, and function of piRNA-producing genes Phillip D. Zamore, P.I. Project Summary In flies, mammals, and worms, PIWI-interacting RNAs (piRNAs) silence transposons and repetitive elements, protecting the germline genome from DNA damage and mutation and ensuring that genetic information passes faithfully from generation to generation. They also have other, poorly understood functions. We propose to define the gene structure, genomic architecture, and regulation of the piRNA-producing genes in mice, flies, and worms. We will then use this information to investigate why some long RNAs make piRNAs while others do not. We will also explore how full-length piRNA precursor transcripts, often tens of kilobases long, are converted into mature piRNAs just 23–35 nt long. Finally, we will test a variety of hypotheses, generated from our data, as to how piRNAs function, particularly in mammals, where most piRNAs do not correspond to transposons. To pursue these aims, we will combine genetics, high-throughput sequencing, and classical biochemistry. We believe that by investigating a common set of questions about the piRNA pathway in three model animals, the conserved and divergent features we discover will yield a deeper understanding of this ancient pathway than could be achieved by working on a single species alone. Relevance Small RNAs called “piRNAs” protect animal sperm and egg precursor cells from damage from selfish genetic elements called transposons or “jumping genes.” Without piRNAs, animals from worms to mammals are sterile, yet we do not understand how piRNAs are made, and for many piRNAs, what they regulate. We hope to obtain a more complete and deeper understanding of this amazing process.