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型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。这些扫描识别出与疾病发病机制和面临传染性病原体、气候变化和饮食限制等历史威胁时的生存相关的基因座，为开发基于基因型的诊断和个性化治疗奠定了基础，以逆转遗传性疾病并预防获得性疾病。然而，很难将致病变异体与它们紧密连锁的附近中性变异体分离。一个潜在的解决方案是测定所有潜在的致病变异的分子功能。然而，单独测定可能与疾病相关的数千种变体是不可行的，特别是鉴于预测大多数负责选择和疾病的变体将是调节性的。我们的目标是阐明新的变异潜在的疾病和适应性表型，通过开发一个高通量筛选转录后调控功能。为了实现这一目标，我们将调整最近描述的大规模平行报告基因测定（MPRA），以允许定量表达和翻译的等位基因差异的数千个变体串联。我们会利用这个 适应MPRA全面测定基因座牵连的疾病和适应的变异与转录后功能。最后，在确定了具有可量化影响的变体之后， 表达或翻译，我们将进行有针对性的后续研究，以获得这些性状背后的转录后过程的机制洞察。这些目标将使我们能够从全基因组扫描疾病和选择，以阐明转录后机制，可以操纵设计新的治疗方法。

 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.