The inert chemical property of RNA modification N6-methyladenosine (m6A) makes it very challenging to detect. Most m6A sequencing methods rely on m6A-antibody immunoprecipitation and cannot distinguish m6A and N6,2'-O-dimethyladenosine modification at the cap +1 position (cap m6Am). Although the two antibody-free methods (m6A-REF-seq/MAZTER-seq and DART-seq) have been developed recently, they are dependent on m6A sequence or cellular transfection. Here, we present an antibody-free, FTO-assisted chemical labeling method termed m6A-SEAL for specific m6A detection. We applied m6A-SEAL to profile m6A landscapes in humans and plants, which displayed the known m6A distribution features in transcriptome. By doing a comparison with all available m6A sequencing methods and specific m6A sites validation by SELECT, we demonstrated that m6A-SEAL has good sensitivity, specificity and reliability for transcriptome-wide detection of m6A. Given its tagging ability and FTO's oxidation property, m6A-SEAL enables many applications such as enrichment, imaging and sequencing to drive future functional studies of m6A and other modifications.

中文翻译：

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用于检测 N6-甲基腺苷的无抗体酶辅助化学方法。

RNA 修饰 N6-甲基腺苷 (m6A) 的惰性化学特性使其检测非常具有挑战性。大多数 m6A 测序方法依赖于 m6A-抗体免疫沉淀，无法区分 m6A 和帽 +1 位（帽 m6Am）处的 N6,2'-O-二甲基腺苷修饰。虽然最近开发了两种无抗体方法（m6A-REF-seq/MAZTER-seq 和 DART-seq），但它们依赖于 m6A 序列或细胞转染。在这里，我们提出了一种无抗体、FTO 辅助的化学标记方法，称为 m6A-SEAL，用于特定的 m6A 检测。我们应用 m6A-SEAL 来分析人类和植物中的 m6A 景观，它显示了转录组中已知的 m6A 分布特征。通过与所有可用的 m6A 测序方法和 SELECT 验证的特定 m6A 位点进行比较，我们证明了 m6A-SEAL 对于 m6A 的全转录组检测具有良好的灵敏度、特异性和可靠性。鉴于其标记能力和 FTO 的氧化特性，m6A-SEAL 能够实现许多应用，例如富集、成像和测序，以推动 m6A 和其他修饰的未来功能研究。