大规模平行报告基因测定:一种分析基因表达调控的新技术

Massively parallel reporter assay:a novel technique for analyzing the regulation of gene expression

大规模平行报告基因测定(massively parallel reporter assay,MPRA)是一种可以同时研究基因组数千个调控元件活性的高通量分析方法。该方法在传统的荧光素酶报告基因载体上引入一段具有唯一标识的条形码,通过二代测序技术对转染前的DNA条形码和转染后的mRNA条形码进行测序,用mRNA和DNA条形码读数的比值来分析顺式调控元件的活性。自MPRA提出以来,已被广泛应用于基因组顺式调控元件和功能性变异的鉴定、转录后调控对表型的影响等方面的研究。本文对MPRA的发展历程、基本原理、实验流程、统计分析方法以及在顺式调控元件和转录后调控方面的应用进行了综述,并对其发展前景进行了展望,以期为相关领域研究人员了解与应用MPRA提供有益参考。

Massively parallel reporter assay(MPRA)is a high-throughput analysis method that can simultaneously investigate the activity of thousands of regulatory elements in the genome.MPRA introduces a uniquely identified barcode on a conventional luciferase reporter gene vector,sequences the DNA barcode before transfection and the mRNA barcode after transfection by next-generation sequencing technology,and uses the ratio of mRNA and DNA barcode reads to analyze the activity of cis-regulatory elements.Since MPRA was proposed,it has been widely used in the identification of genomic cis-regulatory elements and functional variants,the effect of post-transcriptional regulation on phenotypes and so on.In this review,we summarize the development history,basic principles,experimental procedures and statistical analysis methods of MPRA,and its applications in post-transcriptional regulation and cis-regulatory elements.It also provides prospects for its development and useful references for researchers in related fields to understand and apply MPRA.