通过焦磷酸测序法估算5-羟色胺2C受体中五个编辑位点的RNA编辑效率。
Estimating RNA editing efficiency of five editing sites in the serotonin 2C receptor by pyrosequencing.
作者信息
Iwamoto Kazuya, Bundo Miki, Kato Tadafumi
机构信息
Laboratory for Molecular Dynamics of Mental Disorders, Brain Science Institute, RIKEN, Wako-city, Saitama 351-0198, Japan.
出版信息
RNA. 2005 Oct;11(10):1596-603. doi: 10.1261/rna.2114505.
Abstract
Accumulating evidence suggests that altered RNA editing of the serotonin 2C receptor (HTR2C) is involved in the pathophysiology of mental disorders and the action of antidepressants. Estimating RNA editing of HTR2C in various samples is a first step to understanding its pathophysiological roles. Here, we developed a high-throughput quantification method of RNA editing efficiency by pyrosequencing. By optimizing the dispensation order, the RNA editing efficiency of all five RNA editing sites including consecutively ordered sites in HTR2C was obtained. More importantly, our method made it possible to determine the content of partial HTR2C isoforms, which enabled us to monitor possible functional changes of HTR2C. This method was validated in both oligonucleotide and RT-PCR product templates, and showed good correlation with conventional cloning-sequencing analysis. Our method could be a valuable tool in the rapid assessment of RNA editing status, including assessment of natural variations, alterations in disease tissues, and responses to drugs.
摘要
越来越多的证据表明，血清素2C受体（HTR2C）的RNA编辑改变与精神障碍的病理生理学及抗抑郁药的作用有关。估计不同样本中HTR2C的RNA编辑是了解其病理生理作用的第一步。在此，我们开发了一种通过焦磷酸测序对RNA编辑效率进行高通量定量的方法。通过优化加样顺序，获得了HTR2C中包括连续排列位点在内的所有五个RNA编辑位点的RNA编辑效率。更重要的是，我们的方法能够确定部分HTR2C亚型的含量，这使我们能够监测HTR2C可能的功能变化。该方法在寡核苷酸和RT-PCR产物模板中均得到验证，并且与传统的克隆测序分析显示出良好的相关性。我们的方法可能是快速评估RNA编辑状态的有价值工具，包括评估自然变异、疾病组织中的改变以及对药物的反应。
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本文引用的文献
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