Precise Antibody -Independent m6A Identification via 4SedTTP-Involved and FTO-Assisted Strategy at Single-Nucleotide Resolution
Precise Antibody -Independent m6A Identification via 4SedTTP-Involved and FTO-Assisted Strategy at Single-Nucleotide Resolution
复制标题
通过 4SedTTP 参与和 FTO 辅助策略在单核苷酸分辨率下精确鉴定独立于抗体的 m6A
DOI：
10.1021/jacs.7b13633
复制
发表时间：
2018-05-09
影响因子：
15
通讯作者：
Zhou, Xiang
中科院分区：
文献类型：
--
作者：
Hong, Tingting;Yuan, Yushu;Zhou, Xiang
利用创新的检测技术实现m6A在哺乳动物转录组中的精确分布，可以促进我们对其生物学功能的理解。我们特别引入了在脱氧胸苷三磷酸的4位用逐渐增大的原子(硫和硒)取代氧，以削弱其与m6A的碱基对能力，同时保持A-T*碱基对与自然碱基对几乎相同。4SedTTP被证明是一个杰出的候选者，它赋予m6A一个特异的RT截断特征，从而使这种“RT-沉默”的修饰可以在m6A去甲基酶FTO的帮助下通过下一代测序进行检测。这种抗体不依赖、4SedTTP参与和FTO辅助的策略适用于m6A鉴定，甚至适用于单核苷酸分辨未知区域内两个紧密聚集的m6A位点。
Innovative detection techniques to achieve precise m6A distribution within mammalian transcriptome can advance our understanding of its biological functions. We specifically introduced the atom-specific replacement of oxygen with progressively larger atoms (sulfur and selenium) at 4-position of deoxythymidine triphosphate to weaken its ability to base pair with m6A, while maintaining A-T* base pair virtually the same as the natural one. 4SedTTP turned out to be an outstanding candidate that endowed m6A with a specific signature of RT truncation, thereby making this "RT-silent" modification detectable with the assistance of m6A demethylase FTO through next-generation sequencing. This antibody-independent, 4SedTTP-involved and FTO-assisted strategy is applicable in m6A identification, even for two closely gathered m6A sites, within an unknown region at single-nucleotide resolution.