论文标题
利用大规模平行的记者分析进行进化问题
Leveraging massively parallel reporter assays for evolutionary questions
论文作者
论文摘要
进化生物学的一个长期目标是解释基因调节过程如何在物种之间和物种之间有生物多样性。由于从非编码序列预测功能的困难以及非模型分类单元的实验室研究的技术约束,这个问题仍然充满挑战。然而,在功能基因组学的最新方法论发展,即大规模平行的记者测定法(MPRA),使得在单个体外实验中可以测试成千上万的调节活性序列。这样做是通过将传统的,单位的偶发记者测定(例如荧光素酶报告基因测定法)与高通量测序的可扩展性相结合。从这个角度来看,我们讨论了MPRA在进化生物学研究中的执行,优势和局限性。我们回顾了最新的研究,这些研究利用这种方法来解决明确的进化问题,突出了我们认为从MPRA方法中获得的研究设计,这些研究设计特别有利。此外,我们提出了将这些强大测定法扩展到稀有分类单元和具有有限基因组资源的分类单元的解决方案。在此过程中,我们强调了MPRA在非传统模型生物中驱动基因组规模的功能进化遗传学研究的广泛潜力。
A long-standing goal of evolutionary biology is to decode how gene regulatory processes contribute to organismal diversity, both within and between species. This question has remained challenging to answer, due both to the difficulties of predicting function from non-coding sequence, and to the technological constraints of laboratory research with non-model taxa. However, a recent methodological development in functional genomics, the massively parallel reporter assay (MPRA), makes it possible to test thousands to millions of sequences for regulatory activity in a single in vitro experiment. It does so by combining traditional, single-locus episomal reporter assays (e.g., luciferase reporter assays) with the scalability of high-throughput sequencing. In this perspective, we discuss the execution, advantages, and limitations of MPRAs for research in evolutionary biology. We review recent studies that have made use of this approach to address explicitly evolutionary questions, highlighting study designs that we believe are particularly well-positioned to gain from MPRA approaches. Additionally, we propose solutions for extending these powerful assays to rare taxa and those with limited genomic resources. In doing so, we underscore the broad potential of MPRAs to drive genome-scale functional evolutionary genetics studies in non-traditional model organisms.