美国杜克大学医学院的Kate D. Meyer取得一项新进展。研究人员开发了一个名为DART-seq的技术，该方法能够不使用抗体来对全局的m⁶A修饰进行检测。相关论文2019年9月23日在线发表于《自然—方法学》。

研究人员报道了一个名为DART-seq的技术，这是一种无抗体的m⁶A位点检测方法。在DART-seq中，胞苷脱氨酶APOBEC1与m⁶A结合的YTH结构域融合。细胞中APOBEC1-YTH的表达可在与m⁶A残基相邻的位点诱导C-U脱氨，可使用标准RNA序列检测到。 DART-seq可从低至10ng的总RNA中识别出细胞中的数千个m⁶A位点，并可随时间检测细胞中m⁶A的积累。此外，研究人员使用长读本的DART-seq来深入研究了m⁶A沿单个转录本长度的分布。

研究人员表示，m⁶A是一种广泛的RNA修饰，几乎影响信使RNA生命周期的每个方面。全局m⁶A检测方法的发展促进了我们对m⁶A的理解，该方法使用抗体免疫沉淀甲基化的RNA。但是，这些方法有一些局限性，包括对测序RNA要求较高和与其他RNA修饰的交叉反应性。

附：英文原文

Title: DART-seq: an antibody-free method for global m 6 A detection

Author: Kate D. Meyer

Issue&Volume: 2019-09-23

Abstract: 

N6-methyladenosine (m6A) is a widespread RNA modification that influences nearly every aspect of the messenger RNA lifecycle. Our understanding of m6A has been facilitated by the development of global m6A mapping methods, which use antibodies to immunoprecipitate methylated RNA. However, these methods have several limitations, including high input RNA requirements and cross-reactivity to other RNA modifications. Here, we present DART-seq (deamination adjacent to RNA modification targets), an antibody-free method for detecting m6A sites. In DART-seq, the cytidine deaminase APOBEC1 is fused to the m6A-binding YTH domain. APOBEC1-YTH expression in cells induces C-to-U deamination at sites adjacent to m6A residues, which are detected using standard RNA-seq. DART-seq identifies thousands of m6A sites in cells from as little as 10 ng of total RNA and can detect m6A accumulation in cells over time. Additionally, we use long-read DART-seq to gain insights into m6A distribution along the length of individual transcripts.

DOI: 10.1038/s41592-019-0570-0

Source:https://www.nature.com/articles/s41592-019-0570-0