Institute of Pharmacy and Biochemistry, University of Mainz, Mainz, Germany.
RNA Biol. 2010 Mar-Apr;7(2):237-47. doi: 10.4161/rna.7.2.11468. Epub 2010 Mar 8.
RNA nucleotide modifications are typically of low abundance and frequently go unnoticed by standard detection methods of molecular biology and cell biology. With a burst of knowledge intruding from such diverse areas as genomics, structural biology, regulation of gene expression and immunology, it becomes increasingly clear that many exciting functions of nucleotide modifications remain to be explored. It follows in turn that the biology of nucleotide modification and editing is a field poised to rapidly gain importance in a variety of fields. The detection and analysis of nucleotide modifications present a clear limitation in this respect. Here, various methods for detection of nucleotide modifications are discussed based on three discriminating principles, namely physicochemical properties, enzymatic turnover and chemical reactivity. Because the full extent of nucleotide modification across the various RNA species remains ill understood, emphasis is placed on high-throughput techniques with a potential to screen entire transcriptomes.
RNA 核苷酸修饰通常丰度较低,并且经常被分子生物学和细胞生物学的标准检测方法所忽略。随着来自基因组学、结构生物学、基因表达调控和免疫学等不同领域的知识的大量涌现,越来越明显的是,核苷酸修饰的许多令人兴奋的功能仍有待探索。因此,核苷酸修饰和编辑的生物学是一个在许多领域迅速获得重视的领域。核苷酸修饰的检测和分析在这方面存在明显的局限性。在这里,根据理化性质、酶周转率和化学反应性这三个区分原则,讨论了各种检测核苷酸修饰的方法。由于各种 RNA 种类的核苷酸修饰的全貌仍不为人知,因此重点放在具有筛选整个转录组潜力的高通量技术上。