Leveraging massively parallel reporter assays for evolutionary questions

TL;DR

Massively parallel reporter assays (MPRAs) enable high-throughput functional testing of regulatory sequences, offering new opportunities for evolutionary biology research especially in non-model organisms with limited resources.

Contribution

This paper reviews the application of MPRAs in evolutionary biology, highlighting their potential to address long-standing questions about gene regulation and diversity.

Findings

MPRAs can test thousands to millions of sequences simultaneously.

They are particularly useful for studying non-model organisms.

Proposed solutions extend MPRA use to rare taxa with limited genomic data.

Abstract

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.