VARIATION AND REGULATION OF ALTERNATIVE SPLICING IN HUMAN TRANSCRIPTOMES

项目总结 此更新R01项目的中心目标是阐明替代方案的变化和调节 人类转录本中的剪接(AS)。真核细胞产生惊人的调控多样性和复杂性 来自有限一组基因的表型。作为前体，mRNA是产生这一现象的关键机制 监管的多样性。几乎所有人类多外显子基因都是交替拼接的。AS的广泛变化 发生在物种之间、人类种群内，以响应发育信号和环境 微扰，并在疾病发病机制中。尽管AS在基因调控和疾病中的重要性，AS 随着人们对这一领域的广泛兴趣和研究活动，仍有许多悬而未决的问题和重要的 关于人类AS变异的景观、调节和功能后果的知识差距 抄本。用于表征转录组复杂性的强大测序技术(RNA-seq)和 蛋白质-RNA相互作用(CLIP-SEQ)，以及持续存储到公众中的海量数据 为AS的研究创造了令人兴奋和前所未有的机会。该提案整合了数据- 驱动型研究利用大量转录组数据和假设驱动的研究使用分子和 基因组工具。在三个目标中，我们将研究AS在灵长类动物中的进化 (目标1)，遗传变异 和人类群体中AS的表型关联(目标2)，以及整个人类AS的表观遗传调节 组织和细胞状态(目标3)。 我们还将开发和传播创新的计算和统计 使用大型、不同种类的测序数据集进行AS分析的方法。与我们之前的筹资周期一样， 我们将利用我们广泛的专家合作者网络，扩大我们工作的影响，并追求新的 建议项目范围内外的研究机会。总而言之，我们的研究将 对AS的变化、调节和功能产生重要的新颖见解，并广泛创建 用于研究不同生物医学中AS变异和mRNA异构体复杂性的实用计算工具 纪律。

PROJECT SUMMARY The central objective of this renewal R01 project is to elucidate the variation and regulation of alternative splicing (AS) in human transcriptomes. Eukaryotic cells generate astonishing regulatory diversity and complex phenotypes from a finite set of genes. AS of precursor mRNA is a mechanism essential for generating this regulatory diversity. Almost all multi-exon human genes are alternatively spliced. Widespread changes in AS occur between species, within human populations, in response to developmental signals and environmental perturbations, and in disease pathogenesis. Despite the importance of AS in gene regulation and disease, as well as extensive interest and research activities in this field, there remain many open questions and significant knowledge gaps regarding the landscape, regulation, and functional consequence of AS variation in human transcriptomes. Powerful sequencing technologies for characterizing transcriptome complexity (RNA-seq) and protein-RNA interaction (CLIP-seq), as well as the vast amounts of data continuously deposited into the public domain, create exciting and unprecedented opportunities for studies of AS. This proposal integrates data- driven research leveraging big transcriptome data with hypothesis-driven research using molecular and genomic tools. In three aims, we will investigate the evolution of AS in primates (Aim 1), the genetic variation and phenotypic association of AS in human populations (Aim 2), and epigenetic regulation of AS across human tissues and cell states (Aim 3). We will also develop and disseminate innovative computational and statistical methods for analyzing AS using large, heterogeneous sequencing datasets. As in our previous funding cycle, we will tap into our extensive network of expert collaborators to amplify the impact of our work and pursue new research opportunities within and beyond the scope of the proposed project. Collectively, our research will generate significant novel insights into the variation, regulation, and function of AS, and create broadly applicable computational tools for studying AS variation and mRNA isoform complexity in diverse biomedical disciplines.