Vanderbilt University, Department of Pharmacology, 8140 Medical Research Building 3, Nashville, TN 37240-1104, United States.
Missouri State University, Department of Biomedical Sciences, 901 South National Avenue, Springfield, MO 65897, United States.
Methods. 2019 Mar 1;156:40-45. doi: 10.1016/j.ymeth.2018.10.016. Epub 2018 Oct 26.
The deamination of adenosine to inosine by RNA editing is a widespread post-transcriptional process that expands genetic diversity. Selective substitution of inosine for adenosine in pre-mRNA transcripts can alter splicing, mRNA stability, and the amino acid sequence of the encoded protein. The functional consequences of RNA editing-dependent amino acid substitution are known for only a handful of RNA editing substrates. Many of these studies began in heterologous mammalian expression systems; however, the gold-standard for determining the functional significance of transcript-specific re-coding A-to-I editing events is the generation of a mouse model that expresses only one RNA editing-dependent isoform. The frequency of site-specific RNA editing varies spatially, temporally, and in some diseases, therefore, determining the profile of RNA editing frequency is also an important element of research. Here we review the strengths and weaknesses of existing mouse models for the study of RNA editing, as well as methods for quantifying RNA editing frequencies in vivo. Importantly, we highlight opportunities for future RNA editing studies in mice, projecting that improvements in genome editing and high-throughput sequencing technologies will allow the field to excel in coming years.
RNA 编辑将腺苷脱氨为肌苷是一种广泛存在的转录后过程，可扩大遗传多样性。在 mRNA 前体中转录本中肌苷选择性替代腺苷可改变剪接、mRNA 稳定性和编码蛋白的氨基酸序列。RNA 编辑依赖性氨基酸取代的功能后果仅为少数 RNA 编辑底物所知。这些研究中的许多都是从异源哺乳动物表达系统开始的；然而，确定转录特异性重新编码 A 到 I 编辑事件的功能意义的金标准是生成仅表达一种 RNA 编辑依赖性同工型的小鼠模型。特定位置的 RNA 编辑频率在空间上、时间上和在某些疾病中都有所不同，因此，确定 RNA 编辑频率的分布也是研究的一个重要内容。在这里，我们回顾了现有的用于 RNA 编辑研究的小鼠模型的优缺点，以及体内定量 RNA 编辑频率的方法。重要的是，我们强调了未来在小鼠中进行 RNA 编辑研究的机会，预计基因组编辑和高通量测序技术的改进将使该领域在未来几年取得优异成绩。
