Noncoding RNA Laboratory, Department of Molecular Biology and Genetics, İzmir Institute of Technology, Urla, 35430 İzmir, Turkey.
Int J Mol Sci. 2024 Mar 7;25(6):3098. doi: 10.3390/ijms25063098.
Epitranscriptomic mechanisms, which constitute an important layer in post-transcriptional gene regulation, are involved in numerous cellular processes under health and disease such as stem cell development or cancer. Among various such mechanisms, RNA methylation is considered to have vital roles in eukaryotes primarily due to its dynamic and reversible nature. There are numerous RNA methylations that include, but are not limited to, 2'-O-dimethyladenosine (mAm), -methylguanosine (mG), -methyladenosine (mA) and -methyladenosine (mA). These biochemical modifications modulate the fate of RNA by affecting the processes such as translation, target site determination, RNA processing, polyadenylation, splicing, structure, editing and stability. Thus, it is highly important to quantitatively measure the changes in RNA methylation marks to gain insight into cellular processes under health and disease. Although there are complicating challenges in identifying certain methylation marks genome wide, various methods have been developed recently to facilitate the quantitative measurement of methylated RNAs. To this end, the detection methods for RNA methylation can be classified in five categories such as antibody-based, digestion-based, ligation-based, hybridization-based or direct RNA-based methods. In this review, we have aimed to summarize our current understanding of the detection methods for RNA methylation, highlighting their advantages and disadvantages, along with the current challenges in the field.
表观转录组学机制是转录后基因调控的一个重要层面,涉及到健康和疾病状态下的许多细胞过程,如干细胞发育或癌症等。在各种表观转录组学机制中,RNA 甲基化被认为在真核生物中具有重要作用,主要是因为其动态和可逆的性质。有许多种 RNA 甲基化,包括但不限于 2'-O-二甲基腺苷(mAm)、-甲基鸟嘌呤(mG)、-甲基腺苷(mA)和 -甲基腺苷(m6A)。这些生化修饰通过影响翻译、靶标位点确定、RNA 加工、多聚腺苷酸化、剪接、结构、编辑和稳定性等过程来调节 RNA 的命运。因此,定量测量 RNA 甲基化标记的变化对于深入了解健康和疾病状态下的细胞过程非常重要。尽管在全基因组范围内识别某些甲基化标记存在复杂的挑战,但最近已经开发了各种方法来促进甲基化 RNA 的定量测量。为此,RNA 甲基化的检测方法可以分为五类,如基于抗体、基于消化、基于连接、基于杂交或直接基于 RNA 的方法。在这篇综述中,我们旨在总结我们目前对 RNA 甲基化检测方法的理解,强调它们的优缺点,以及该领域目前面临的挑战。