芝加哥大学的Bryan C. Dickinson等报道了一项人工进化逆转录酶技术，并将其用于筛选可检测人类信使RNA中N1-甲基腺苷修饰位点的逆转录酶。

研究描述了一种用于定向进化的通用平台，其可快速选择逆转录酶，该逆转录酶在逆转录过程中在给定类型的RNA修饰的位点加上突变，从而允许位点特异性识别修饰。为了开发和验证该平台，研究人员人工进化了针对N1-甲基腺苷（m1A）的HIV-1逆转录酶。反复数次的筛选得到了逆转录酶，其在m1A位点具有强大的识别性和高突变率。最佳进化逆转录酶能够检测出特征明确的m1A位点，并揭示了人类mRNA中的数百个m1A位点。

这项工作开发并验证了该逆转录酶进化平台，并为m1A生物学的研究提供了新的工具、分析方法和数据集。

研究表示，对信使RNA的化学修饰越来越被认为是遗传信息流中的关键调控层面，但目前缺乏以全转录组和位点特异性方式监测RNA修饰的定量工具。

附：

Title: Evolution of a reverse transcriptase to map N 1 -methyladenosine in human messenger RNA

Author: Huiqing Zhou, Simone Rauch, Qing Dai, Xiaolong Cui, Zijie Zhang, Sigrid Nachtergaele, Caraline Sepich, Chuan He, Bryan C. Dickinson

Issue&Volume: 2019-09-23

Abstract: 

Chemical modifications to messenger RNA are increasingly recognized as a critical regulatory layer in the flow of genetic information, but quantitative tools to monitor RNA modifications in a whole-transcriptome and site-specific manner are lacking. Here we describe a versatile platform for directed evolution that rapidly selects for reverse transcriptases that install mutations at sites of a given type of RNA modification during reverse transcription, allowing for site-specific identification of the modification. To develop and validate the platform, we evolved the HIV-1 reverse transcriptase against N1-methyladenosine (m1A). Iterative rounds of selection yielded reverse transcriptases with both robust read-through and high mutation rates at m1A sites. The optimal evolved reverse transcriptase enabled detection of well-characterized m1A sites and revealed hundreds of m1A sites in human mRNA. This work develops and validates the reverse transcriptase evolution platform, and provides new tools, analysis methods and datasets to study m1A biology.

DOI: 10.1038/s41592-019-0550-4

Source:https://www.nature.com/articles/s41592-019-0550-4