N6-甲基腺嘌呤(m6A)是真核生物RNA分子中最常见及丰度最高的修饰方式,m6A占所有RNA碱基修饰的比例超过80%,而且在不同的物种中存在。m6A主要分布在mRNA中,同时也存在于非编码RNA中。mRNA转录本中的m6A相对丰度已经被证实能够影响RNA代谢过程,如剪切、出核转运、翻译能力和稳定性、RNA转录等。异常的m6A甲基化水平受到m6A RNA甲基化酶和去甲基化酶的影响,从而导致RNA的异常功能及疾病的发生。因此,RNA中动态可逆的m6A化学修饰能够作为新的表观遗传学标志物,具有重要的生物学作用。认识RNA转录本中m6A甲基化水平和分布能够有利于疾病的诊断和治疗。
本检测采用EpiQuik m6A RNA甲基化定量检测试剂盒,通过比色的方法定量总RNA中m6A 的甲基化程度,适用于哺乳动物、植物、真菌、细菌和病毒等物种。检测原理是在特定的结合缓冲液中,将待检测的total RNA或者标准品结合到微孔板表面,然后依次加入特异性的m6A捕获抗体、检测抗体和显色试剂,最终显示的颜色深浅与样品中m6A的浓度呈正比,利用酶标仪在450nm波长下测定吸光度(OD值),通过标准曲线即可计算样品中m6A的绝对含量,同时通过total RNA总量校正可对不同样本中m6A含量进行比较。
1. Slobodin B et. al. (April 2017). Transcription Impacts the Efficiency of mRNA Translation via Co-transcriptional N6-adenosine Methylation. Cell. 169(2):326-337.e12.
2. Elkashef SM et. al. (May 2017). IDH Mutation, Competitive Inhibition of FTO, and RNA Methylation. Cancer Cell. 31(5):619-620.
3. Song H et. al. (March 2019). METTL3 and ALKBH5 oppositely regulate m6A modification of TFEB mRNA, which dictates the fate of hypoxia/reoxygenation-treated cardiomyocytes. Autophagy. :1-19.
4. Panneerdoss S et. al. (October 2018). Cross-talk among writers, readers, and erasers of m6A regulates cancer growth and progression. Sci Adv. 4(10):eaar8263.
5. Zheng ZQ et. al. (July 2019). Long non-coding RNA FAM225A promotes nasopharyngeal carcinoma tumorigenesis and metastasis by acting as ceRNA to sponge miR-590-3p/miR-1275 and upregulate ITGB3. Cancer Res.
6. Niu Y et. al. (March 2019). RNA N6-methyladenosine demethylase FTO promotes breast tumor progression through inhibiting BNIP3. Mol Cancer. 18(1):46.