Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
Biotechniques. 2012 Feb;52(2):109-11. doi: 10.2144/000113801.
Oxidative RNA damage has been linked to loss of RNA function and to the development of many human age-related diseases. Consequently, a need exists for methods to identify and quantify the extent of RNA oxidation on a genome-wide basis. We developed such a method by combining affinity selection of mRNA containing 8-hydroxyguanine with high throughput DNA sequencing. We demonstrate that this assay is suitable for detecting differences in the extent of oxidation between RNA transcripts. We applied this method to the yeast Saccharomyces cerevisiae grown under physiological conditions and in response to hydrogen peroxide, and detected significantly oxidized RNA transcripts.
氧化的 RNA 损伤与 RNA 功能丧失以及许多与年龄相关的人类疾病的发展有关。因此，需要开发在全基因组范围内识别和量化 RNA 氧化程度的方法。我们通过结合含有 8-羟基鸟嘌呤的 mRNA 的亲和选择与高通量 DNA 测序来开发这种方法。我们证明该测定法适用于检测 RNA 转录本之间氧化程度的差异。我们将该方法应用于在生理条件下和响应于过氧化氢生长的酵母 Saccharomyces cerevisiae 中，并检测到明显氧化的 RNA 转录本。
