A versatile method to design stem-loop primer-based quantitative PCR assays for detecting small regulatory RNA molecules
| Autor: | Attila Varga, Balazs Dezso, Attila Horvath, Zoltan Simandi, Éva Várallyay, Julianna Hulvely, Zoltán Havelda, Laszlo Nagy, Erzsébet Szabó, Balint Domokos, Zsolt Török, Balint L. Balint, Zsolt Czimmerer, Maria Balogh |
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| Rok vydání: | 2012 |
| Předmět: |
Small interfering RNA
Science Computational biology Biology Regulatory Sequences Ribonucleic Acid Polymerase Chain Reaction Biochemistry Cell Line Small hairpin RNA Mice RNA interference Molecular cell biology Transduction Genetic microRNA Gene expression Gene Knockdown Techniques Genetics Animals Humans Elméleti orvostudományok RNA Small Interfering DNA Primers Multidisciplinary Base Sequence Inverted Repeat Sequences Lentivirus RNA Orvostudományok RNA stability Stem-loop Nucleic acids RNA processing Regulatory sequence Medicine Epigenetics Research Article |
| Zdroj: | PLoS ONE PLoS ONE, Vol 8, Iss 1, p e55168 (2013) |
| ISSN: | 1932-6203 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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