Department of Plant Ecophysiology, Institute of Experimental Biology, Faculty of Biology, School of Natural Sciences, Adam Mickiewicz University, Poznan, Poland.
Methods Mol Biol. 2022;2526:125-134. doi: 10.1007/978-1-0716-2469-2_9.
Oxidation of RNA is associated with the development of numerous disorders including Alzheimer's and Parkinson's diseases, amyotrophic lateral sclerosis (ALS), cancer, and diabetes. Additionally, a correlation has been found between increase in RNA oxidation and the process of aging. In plants, elevated level of oxidatively modified transcripts has been detected during alleviation of seeds dormancy and stress response. Increasing interest on the topic of RNA oxidative modifications requires elaboration of new laboratory techniques. So far, the most common method used for the assessment of RNA oxidation is quantification of 8-hydroxyguanine (8-OHG). However, reactive oxygen species (ROS) induce also numerous other changes in nucleic acids, including formation of abasic sites (AP-sites). Recently, the level of AP-sites in RNA has been measured with the use Aldehyde Reactive Probe (ARP). In the present chapter, we describe application of this technique for the evaluation of the level of AP-sites in plant transcripts.
RNA 的氧化与许多疾病的发展有关，包括阿尔茨海默病和帕金森病、肌萎缩侧索硬化症 (ALS)、癌症和糖尿病。此外，还发现 RNA 氧化的增加与衰老过程有关。在植物中，在缓解种子休眠和应对压力时，检测到氧化修饰转录本水平升高。对 RNA 氧化修饰这一课题的兴趣日益增加，这需要开发新的实验室技术。到目前为止，评估 RNA 氧化的最常用方法是定量检测 8-羟基鸟嘌呤 (8-OHG)。然而，活性氧 (ROS) 还会引起核酸中许多其他变化，包括形成无碱基位点 (AP 位点)。最近，使用醛反应探针 (ARP) 测量了 RNA 中 AP 位点的水平。在本章中，我们描述了该技术在评估植物转录本中 AP 位点水平的应用。
