Genome-wide validation of posttranscriptional variation in selection and disease

 描述(由申请人提供)：来自疾病队列和不同种族群体的基因数据的涌入显着增加了全基因组扫描对疾病关联和阳性选择的能力。这些扫描在面临传染病、气候变化和饮食限制等历史威胁的情况下识别与疾病发病机制和生存相关的基因座，为基于基因的诊断和个性化治疗的发展奠定了基础，以逆转遗传病和预防获得性疾病。然而，很难将致病变种从与它们密切相关的附近的中性变种中分离出来。一种可能的解决方案是分析分子功能的所有潜在因果变量。然而，单独分析可能与疾病相关的数千个变异是不可行的，特别是考虑到预测大多数负责选择和疾病的变异将是受调控的。我们的目标是通过开发高通量的转录后调节功能筛查来阐明疾病和适应性表型的新变种。为了实现这一目标，我们将对最近描述的大规模并行报告程序分析(MPRA)进行改造，以允许对数千个变体的表达和翻译中的等位基因差异进行量化。我们将利用这一点 采用MPRA来全面分析与疾病有关的基因座，以及对具有转录后功能的变体的适应性。最后，在确定了具有量化影响的变体之后 对于表达或翻译，我们将进行有针对性的后续研究，以获得对这些特征潜在的转录后过程的机械性洞察。这些目标将使我们能够从全基因组扫描疾病和选择转移到阐明转录后机制，这些机制可以被操纵来设计新的治疗方法。

 DESCRIPTION (provided by applicant): An influx of genotype data from disease cohorts and diverse ethnic groups is markedly increasing the power of genome-wide scans for disease association and positive selection. These scans identify loci relevant to the pathogenesis of disease and survival in the face of historic threats such as infectious agents, climate change, and dietary limitations, setting the stage for the development of genotype-based diagnostics and personalized therapeutics to reverse inherited disease and protect against acquired disease. However, it is difficult to isolate disease-causing variants from nearby neutral variants to which they are closely linked. One potential solution is to assay all potentially causal variants for molecular function. However, it is infeasible to individually assay the thousands of variants potentially associated with disease, especially in light of the prediction that most variants responsible for selection and disease will be regulatory. We aim to elucidate novel variants underlying disease and adaptive phenotypes by developing a high-throughput screen for posttranscriptional regulatory function. To accomplish this goal, we will adapt the recently described Massively Parallel Reporter Assay (MPRA) to allow the quantification of allelic differences in expression and translation for thousands of variants in tandem. We will utilize this adapted MPRA to comprehensively assay loci implicated in disease and adaptation for variants with posttranscriptional function. Finally, after identifying variants with quantifiable effects on expression or translation, we will perform targeted follow-up studies to gain mechanistic insight into posttranscriptional processes underlying these traits. These aims will enable us to move from genome-wide scans for disease and selection to elucidation of posttranscriptional mechanisms that may be manipulated to devise novel therapeutic approaches.