北京大学伊成器研究组揭示了温和和超快GLORI能够从低输入样品中绝对定量m⁶A甲基组。这一研究成果于2025年5月5日发表在国际顶尖学术期刊《自然—方法学》上。

研究团队之前报道过GLORI，这是一种化学辅助方法，可实现无偏和精确的m⁶A测量。

然而，其漫长的反应时间和严重的RNA降解限制了其适用性，特别是对于低输入的样品。小组提出了两种更新的GLORI方法，它们超快，温和，能够比RNA起始材料少一到两个数量级的绝对m⁶A定量：GLORI 2.0与来自约10,000个细胞的RNA兼容，提高了转录组范围和位点特异性m⁶A检测的灵敏度；GLORI 3.0进一步利用逆转录沉默载体RNA，实现低至500-1,000个细胞的m⁶A定量。使用来自海马背侧的有限RNA，课题组揭示了突触相关基因集的高修饰水平。小组设想更新后的GLORI方法将极大地扩展m⁶A绝对定量在生物学中的适用性。

据悉，N⁶-甲基腺苷（m⁶A）的绝对定量方法已成为表观转录组学研究的有力工具。

附：英文原文

Title: Mild and ultrafast GLORI enables absolute quantification of m6A methylome from low-input samples

Author: Sun, Hanxiao, Lu, Bo, Zhang, Zeyu, Xiao, Ye, Zhou, Zhe, Xi, Lin, Li, Zhichao, Jiang, Zhe, Zhang, Jiayi, Wang, Meng, Liu, Cong, Ma, Yichen, Peng, Jinying, Wang, Xiu-Jie, Yi, Chengqi

Issue&Volume: 2025-05-05

Abstract: Methods for absolute quantification of N6-methyladenosine (m6A) have emerged as powerful tools in epitranscriptomics. We previously reported GLORI, a chemical-assisted approach to achieve unbiased and precise m6A measurement. However, its lengthy reaction time and severe RNA degradation have limited its applicability, particularly for low-input samples. Here, we present two updated GLORI approaches that are ultrafast, mild and enable absolute m6A quantification from one to two orders of magnitude less than the RNA starting material: GLORI 2.0 is compatible with RNA from ~10,000 cells and enhances sensitivity for both transcriptome-wide and locus-specific m6A detection; GLORI 3.0 further utilizes a reverse transcription-silent carrier RNA to achieve m6A quantification from as low as 500–1,000 cells. Using limited RNA from mouse dorsal hippocampus, we reveal a high modification level in synapse-related gene sets. We envision that the updated GLORI methods will greatly expand the applicability of absolute quantification of m6A in biology.

DOI: 10.1038/s41592-025-02680-9

Source: https://www.nature.com/articles/s41592-025-02680-9