研究人员描绘哺乳动物转录组中m6ARNA的修饰图谱
2022-03-15   阅读:575   来源:自然

美国芝加哥大学何川、Mengjie Chen、Lulu Hu和贝克曼研究所Jianjun Chen研究组,在单碱基分辨率水平测量了哺乳动物转录组中N6-甲基腺苷(m6A) RNA修饰位点。该项研究成果发表在2022年3月14日出版的《自然-生物技术》上。

为了研究RNA m6A修饰的功能,研究人员研发了m6A选择性烯丙基化学标记和测序(m6A-SAC-seq),这是一种以单核苷酸分辨率对m6A在全转录组进行定量的作图方法。该方法仅需要约30 ng的poly (A)或rRNA缺失RNA。研究人员绘制了细胞系RNA和人类造血干细胞和祖细胞(HSPC)体外单核细胞生成过程中m6A修饰的化学计量。

研究确定了许多细胞状态特异性m6A位点,其甲基化状态在细胞分化过程中是高度动态的。研究还发现m6A化学计量以及转录水平的变化可能受对HSPC分化至关重要的关键转录因子(TF) 的调节。m6A-SAC-se是一种高效的定量方法,仅使用有限的RNA来揭示m6A位点在不同生物过程中的动力学和功能。

据了解,因无法在整个转录组中绘制单个m6A修饰位点,此前,RNA m6A修饰的功能研究受到阻碍。

附:英文原文

Title: m6A RNA modifications are measured at single-base resolution across the mammalian transcriptome

Author: Hu, Lulu, Liu, Shun, Peng, Yong, Ge, Ruiqi, Su, Rui, Senevirathne, Chamara, Harada, Bryan T., Dai, Qing, Wei, Jiangbo, Zhang, Lisheng, Hao, Ziyang, Luo, Liangzhi, Wang, Huanyu, Wang, Yuru, Luo, Minkui, Chen, Mengjie, Chen, Jianjun, He, Chuan

Issue&Volume: 2022-03-14

Abstract: Functional studies of the RNA N6-methyladenosine (m6A) modification have been limited by an inability to map individual m6A-modified sites in whole transcriptomes. To enable such studies, here, we introduce m6A-selective allyl chemical labeling and sequencing (m6A-SAC-seq), a method for quantitative, whole-transcriptome mapping of m6A at single-nucleotide resolution. The method requires only ~30ng of poly(A) or rRNA-depleted RNA. We mapped m6A modification stoichiometries in RNA from cell lines and during in vitro monocytopoiesis from human hematopoietic stem and progenitor cells (HSPCs). We identified numerous cell-state-specific m6A sites whose methylation status was highly dynamic during cell differentiation. We observed changes of m6A stoichiometry as well as expression levels of transcripts encoding or regulated by key transcriptional factors (TFs) critical for HSPC differentiation. m6A-SAC-seq is a quantitative method to dissect the dynamics and functional roles of m6A sites in diverse biological processes using limited input RNA.

DOI: 10.1038/s41587-022-01243-z

编辑:小柯机器人

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