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新方法实现强大的m6A定量分析
作者:小柯机器人 发布时间:2021/9/8 16:41:37

以色列魏茨曼科学研究所Schraga Schwartz团队开发出新方法实现强大的N6-甲基腺苷(m6A)定量分析。相关论文于2021年9月3日在线发表在《自然—方法学》杂志上。

研究人员开发了m6A-seq2,依靠条形码和集合样品的多重m6A免疫沉淀。m6A-seq2可以大大增加产量,同时降低技术变异性、输入材料的要求和成本。最后,研究人员为m6A的基因级定量开发了一种计算方法。研究人员证明,这一指标可以解释小鼠胚胎干细胞中RNA半衰期的大约30%改变,从而确定m6A是RNA稳定性的一个主要驱动因素。

据了解,m6A是哺乳动物中最普遍的信使RNA的修饰。为了鉴定其功能和动态,目前迫切需要在三个层面上对m6A进行量化:位点、基因和样本。目前的方法只能有限地满足这些需求。

附:英文原文

Title: Multiplexed profiling facilitates robust m6A quantification at site, gene and sample resolution

Author: Dierks, David, Garcia-Campos, Miguel Angel, Uzonyi, Anna, Safra, Modi, Edelheit, Sarit, Rossi, Alice, Sideri, Theodora, Varier, Radhika A., Brandis, Alexander, Stelzer, Yonatan, van Werven, Folkert, Scherz-Shouval, Ruth, Schwartz, Schraga

Issue&Volume: 2021-09-03

Abstract: N6-methyladenosine (m6A) is the most prevalent modification of messenger RNA in mammals. To interrogate its functions and dynamics, there is a critical need to quantify m6A at three levels: site, gene and sample. Current approaches address these needs in a limited manner. Here we develop m6A-seq2, relying on multiplexed m6A-immunoprecipitation of barcoded and pooled samples. m6A-seq2 allows a big increase in throughput while reducing technical variability, requirements of input material and cost. m6A-seq2 is furthermore uniquely capable of providing sample-level relative quantitations of m6A, serving as an orthogonal alternative to mass spectrometry-based approaches. Finally, we develop a computational approach for gene-level quantitation of m6A. We demonstrate that using this metric, roughly 30% of the variability in RNA half life in mouse embryonic stem cells can be explained, establishing m6A as a main driver of RNA stability. m6A-seq2 thus provides an experimental and analytic framework for dissecting m6A-mediated regulation at three different levels. This work describes m6A-seq2, which utilizes multiplexed N6-methyladenosine (m6A) immunoprecipitation of pre-barcoded and pooled samples to reduce the technical variability and allows direct comparison of m6A at the site, gene and sample level.

DOI: 10.1038/s41592-021-01242-z

Source: https://www.nature.com/articles/s41592-021-01242-z

期刊信息

Nature Methods:《自然—方法学》,创刊于2004年。隶属于施普林格·自然出版集团,最新IF:28.467
官方网址:https://www.nature.com/nmeth/
投稿链接:https://mts-nmeth.nature.com/cgi-bin/main.plex