Single molecule detection of L1 insertions and intermediates

项目摘要 转座基因是人类可遗传基因变异的主要来源，可通过以下途径导致疾病 导致体细胞突变。该应用程序专注于基因组工具和技术的开发，以 评估这些转座子对正常组织中的体细胞嵌合体的贡献。具体来说，我们将 开发工具和技术来检测长分布的Element-1(Line-1，L1)序列中的变异。L1是 人类唯一活跃的自我繁殖蛋白质编码转座子。我们每个人都继承了不同的 活跃的L1基因座，每一个典型地通过其内部启动子的甲基化而被抑制。在这里，我们建议 开发单细胞映射方法来检测从头L1插入，并将这些与长阅读相结合 测序方法，以揭示源元素的表观遗传状态。我们还将开创高度创新的局面 检测靶引发逆转录(TPRT)反应中间产物的方法 L1插入事件。我们的多学科团队汇集了具有反转录转座子专业知识的研究人员 生物学和遗传变异；表观遗传学领域的创新；以及分子生物学和 计算基因组学方法的发展。我们在UG3/UH3项目中的共同目标是提供支持 人类组织间体细胞镶嵌(SMAHTT)网络的工具和技术。

Project Summary Transposable elements are major sources of heritable human genetic variation and can contribute to disease by causing somatic mutations. This application focuses on the development of genomic tools and technologies to assess the contributions of these transposons to somatic mosaicism in normal tissues. Specifically, we will develop tools and technologies to detect variants in long interspersed element-1 (LINE-1, L1) sequences. L1 is the only active self-propagating protein-coding transposon in humans. We each inherit a distinct complement of active L1 loci, each of which is typically repressed through methylation of its internal promoter. Here, we propose to develop single cell mapping approaches to detect de novo L1 insertions and integrate these with long-read sequencing approaches to reveal the epigenetic status of source elements. We will also pioneer highly innovative approaches to detect intermediates of the target primed reverse transcription (TPRT) reaction that gives rise to L1 insertion events. Our multidisciplinary team brings together investigators with expertise in retrotransposon biology and genetic variation; innovation in the field of epigenetics; and experience with molecular biology and computational genomics methods development. Our shared goal in this UG3/UH3 project is to provide enabling tools and technologies to the Somatic Mosaicism across Human Tissues (SMaHT) Network.