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A LINE-1 Retrotransposition-Deficient Mouse Genome

LINE-1 逆转录转座缺陷小鼠基因组

基本信息

批准号：
9925240
负责人：
ALEX BORTVIN
金额：
$ 21.47万
依托单位：
CARNEGIE INSTITUTION OF WASHINGTON, D.C.
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-02 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9925240
关键词：
Address Aging Autoimmune Diseases Biological Assay CRISPR/Cas technology Cell Culture Techniques Clone Cells Collaborations Complement Complex Correlative Study Cytidine Deaminase Cytosine DNA Sequence DNA Transposable Elements Development Disease Elements Embryology Embryonic Development Evolution Foundations Future Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genome Genomics Goals Guide RNA Health Human Institution Introns L1 Elements Length Long Interspersed Elements Malignant Neoplasms Mediating Messenger RNA Monitor Mus Mutagenesis Mutate Nonsense Codon Oncology Oogenesis Pathologic Processes Pathology Phenotype Problem Solving Process Production Proteins RNA-Binding Proteins RNA-Directed DNA Polymerase Rattus Reporter Repression Research Residual state Resources Retrotransposition Retrotransposon Role Science Site Structure Testing Tetanus Helper Peptide Tetracyclines Tissues Transgenes Universities Untranslated RNA Untranslated Regions Uracil base embryonic stem cell endonuclease human disease in vivo in vivo evaluation inhibitor/antagonist interest mammalian genome medical schools mouse genome neurogenesis next generation sequencing prevent stem cell model transposon/insertion element vertebrate genome

项目摘要

Project Summary Transposable elements (TEs) are discrete mobile DNA sequences whose intact and mutated copies constitute a substantial fraction of host genomes. Over the past decade, numerous studies implicated TEs in the evolution of genomes and gene expression networks. However, whether TEs contribute to development or disease is an open crucial question answering which might expand and even radically change our views on the genetic foundations of human health and pathology. Of particular interest to human health is a Long Interspersed Element 1 (LINE-1 or L1). For example, over a hundred single-gene human diseases arose from L1 insertions. Intriguingly, recent studies implicated L1 retrotransposition in normal development (embryogenesis, oogenesis, neurogenesis), aging and disease (autoimmune disorders, cancers). However, despite a multitude of correlative studies, none have established a causative role of L1 retrotransposition in these processes to date. The primary challenge of studying endogenous L1s lies in the large numbers of full-length intact L1s in mammalian genomes. Experimental reduction of TE activity at the transcriptional or post-transcriptional levels does not provide specific, stable and long-term L1 repression in complex tissues. This R21 proposal aims to make the first step toward closing this gap in our understanding of the contribution of L1 to development and disease. We propose an experimental strategy to gradually and systematically inactivate full-length L1 elements in the mouse genome using the embryonic stem cell (ESC) model. We will monitor and quantify the extent of L1 mutagenesis and ultimately identify candidate ESC clones whose developmental potential we will subsequently test in vivo. The proposed research will provide the foundation for future rigorous tests for the role of L1s in development and disease. Relevance Studies over the past decade implicated mobile element retrotransposon L1 in healthy development and disease. If true, this contention will radically expand our understanding of the genetic foundations of human health and pathology. We propose to test this hypothesis by systematically inactivating L1 elements in the genome of mouse embryonic stem cells and characterizing the developmental potential of genomes lacking active L1 elements.

项目概要转座元件 (TE) 是离散的可移动 DNA 序列，其完整和突变的拷贝构成了宿主基因组的很大一部分。在过去的十年中，大量研究表明 TE 与进化有关基因组和基因表达网络。然而，TE 是否会导致发育或疾病是一个问题开放的关键问题回答可能会扩大甚至从根本上改变我们对遗传的看法人类健康和病理学的基础。对人类健康特别感兴趣的是长期散布的元素 1（LINE-1 或 L1）。例如，超过 100 种单基因人类疾病是由 L1 插入引起的。有趣的是，最近的研究表明 L1 逆转录转座参与正常发育（胚胎发生、卵子发生、神经发生）、衰老和疾病（自身免疫性疾病、癌症）。然而，尽管存在大量相关的迄今为止，尚无研究证实 L1 逆转录转座在这些过程中起因果作用。初级研究内源性 L1 的挑战在于哺乳动物基因组中存在大量全长完整的 L1。实验性降低转录或转录后水平的 TE 活性并不能提供复杂组织中特异性、稳定和长期的 L1 抑制。 R21提案旨在迈出第一步旨在缩小我们对 L1 对发育和疾病的贡献的理解上的差距。我们建议一种逐步、系统地灭活小鼠基因组中全长 L1 元件的实验策略使用胚胎干细胞（ESC）模型。我们将监测和量化 L1 突变的程度最终确定候选ESC克隆，我们随后将在体内测试其发育潜力。这拟议的研究将为未来 L1 在发育和发育中的作用进行严格测试奠定基础。疾病。关联过去十年的研究表明，移动元件逆转录转座子 L1 与健康发育和疾病有关。如果属实，这一论点将从根本上扩展我们对人类健康和遗传基础的理解。病理。我们建议通过系统地灭活基因组中的 L1 元件来检验这一假设。小鼠胚胎干细胞并表征缺乏活性 L1 的基因组的发育潜力元素。