A versatile method to design stem-loop primer-based quantitative PCR assays for detecting small regulatory RNA molecules.
A versatile method to design stem-loop primer-based quantitative PCR assays for detecting small regulatory RNA molecules.
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一种用于设计基于茎环的定量PCR测定的多功能方法，用于检测小调节RNA分子。
DOI：
10.1371/journal.pone.0055168
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Balint BL
中科院分区：
文献类型：
--
作者：
Czimmerer Z;Hulvely J;Simandi Z;Varallyay E;Havelda Z;Szabo E;Varga A;Dezso B;Balogh M;Horvath A;Domokos B;Torok Z;Nagy L;Balint BL
短调控RNA已被鉴定为真核生物中基因表达的关键调控因子。它们参与生理和病理过程的调节，如胚胎发育、免疫调节和癌症。它们的相关特征之一是它们的高稳定性，这使它们成为用作生物标志物的优秀候选者。它们的数量不断增加，因为下一代测序方法揭示了它们合成的越来越多的细节。这些新的发现旨在为单个短调控RNA提供新的检测方法，以便能够确认原始数据并表征不同生物条件下新鉴定的亚型。我们开发了一种灵活的方法来设计非常灵敏和稳健的RT-qPCR检测。新设计的检测方法在植物、小鼠甚至人类福尔马林固定石蜡包埋组织的样品中进行了广泛的测试。此外，我们已经表明，这些测定能够定量内源性产生的shRNA分子。任何希望使用稳健灵活的系统定量分析成熟调控RNA的人都可以免费获得该检测设计方法。
Short regulatory RNA-s have been identified as key regulators of gene expression in eukaryotes. They have been involved in the regulation of both physiological and pathological processes such as embryonal development, immunoregulation and cancer. One of their relevant characteristics is their high stability, which makes them excellent candidates for use as biomarkers. Their number is constantly increasing as next generation sequencing methods reveal more and more details of their synthesis. These novel findings aim for new detection methods for the individual short regulatory RNA-s in order to be able to confirm the primary data and characterize newly identified subtypes in different biological conditions. We have developed a flexible method to design RT-qPCR assays that are very sensitive and robust. The newly designed assays were tested extensively in samples from plant, mouse and even human formalin fixed paraffin embedded tissues. Moreover, we have shown that these assays are able to quantify endogenously generated shRNA molecules. The assay design method is freely available for anyone who wishes to use a robust and flexible system for the quantitative analysis of matured regulatory RNA-s.
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影响因子：
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