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

项目三.了解piRNA产生基因的结构、调控和功能 菲利普D.扎莫尔私家侦探 项目摘要 在苍蝇、哺乳动物和蠕虫中，PIWI相互作用RNA（piRNA）沉默转座子和重复元件， 保护生殖系基因组免受DNA损伤和突变，并确保遗传信息通过 一代又一代的忠实。它们还有其他鲜为人知的功能。我们建议 定义基因结构，基因组结构，以及小鼠，苍蝇， 还有蠕虫然后，我们将使用这些信息来研究为什么一些长RNA会产生piRNA，而另一些长RNA会产生piRNA。 不要。我们还将探索全长皮尔纳前体转录物（通常为数十个碱基长）是如何在细胞中表达的。 转化为仅23-35 nt长的成熟piRNA。最后，我们将测试各种假设，从 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，我们将联合收割机结合遗传学、高通量测序和经典的 生物化学我们认为，通过研究三个国家中关于皮尔纳途径的一组常见问题， 模式动物，我们发现的保守和分歧的特点将产生更深入的了解这一点 这是一个古老的途径，而不是仅仅通过研究单个物种来实现。 相关性 被称为“piRNA”的小RNA保护动物精子和卵子前体细胞免受自私遗传物质的损害。 转座子或“跳跃基因”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piRNA是如何产生的，对于许多piRNA，它们调节什么。我们希望获得 更完整更深入地了解这个惊人的过程

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.