Abstract：
Recent advancements in detection and mapping methods have enabled researchers to uncover the biological importance of RNA chemical modifications, which play a vital role in post-transcriptional gene regulation. Although numerous types of RNA modifications have been identified in higher eukaryotes, only a few have been extensively studied for their biological functions. Of these, N6-methyladenosine (m6A) is the most prevalent and important mRNA modification that influences various aspects of RNA metabolism, including mRNA stability, degradation, splicing, alternative polyadenylation, export, and localization, as well as translation. Thus, they have implications for a variety of biological processes, including growth, development, and stress responses. The m6A deposition or removal on transcripts is dynamic and is altered in response to internal and external cues. Because this mark can alter gene expression under stress conditions, it is essential to identify the transcripts that can acquire or lose this epitranscriptomic mark upon exposure to stress conditions. Here we describe a step-by-step protocol for identifying stress-responsive transcriptome-wide m6A changes using RNA immunoprecipitation followed by high-throughput sequencing (MeRIP-seq).
摘要：
检测和作图方法的最新进展使研究人员能够揭示RNA化学修饰的生物学重要性。在转录后基因调控中起着至关重要的作用。尽管在高等真核生物中已经鉴定出许多类型的RNA修饰,只有少数人对其生物学功能进行了广泛研究。其中,N6-甲基腺苷（m6A）是影响RNA代谢各个方面的最普遍和重要的mRNA修饰。包括mRNA稳定性，降解，拼接,选择性聚腺苷酸化，export,和本地化，以及翻译。因此，它们对各种生物过程有影响，包括增长,发展,和应激反应。转录物上的m6A沉积或去除是动态的并且响应于内部和外部提示而改变。因为这个标记可以在胁迫条件下改变基因表达，在暴露于应激条件时，确定可以获得或失去这种表观基因组标记的转录本是至关重要的。在这里，我们描述了使用RNA免疫沉淀，然后进行高通量测序（MeRIP-seq）来识别应激反应性转录组m6A变化的逐步方案。
