Pitfalls and problems of miRNA expression analysis
Presenter: Michela Denti PhD, University of Trento, Italy
Prof. Denti has an unavoidable commitment that came up on short notice and has arranged for Simone Detassis from her lab to give the webinar in her place.
Date: February 12, 2019
microRNAs are intensively studied as tools for personalized medicine because they encompass many ideal characteristics of biomarkers.
However, some problems, which I will discuss, still limit the use of microRNAs in personalized medicine.
- The source of microRNAs and the extraction method: the influence on recovery and final outcome may be substantial, especially for RNA extracted from biofluids.
- The detection method: the short sequence of microRNAs impedes an easy design of probes, limiting also the discrimination between pri-, pre- and mature forms. Moreover, despite the ease of use and accessibility of qRT-PCR, ddPCR, microarrays and NGS as main detection techniques for nucleic acids, they lack strong sensibility and accuracy, especially at single base resolution.
- The normalization of the signal. In fact, as the analysis is about a relative expression, the choice of a good normalizer is fundamental and challenging Several authors have concluded that a universal endogenous control is unlikely to be discovered and a suitable reference should be assessed every time considering the different biological conditions of the samples.
The presentation will be followed by a Q&A session. A recording of the webinar will be made available on this page.
Recording of the webinar: Click play to view
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Prof. Michela Alessandra Denti is Principal Investigator of the Laboratory of RNA Biology and Biotechnology at the Centre for Integrative Biology of the University of Trento, Italy since 2008. Her laboratory has two main research interests: modulation of RNA splicing as a cure for inherited diseases and microRNAs as biomarkers of cancer, cardiac and neurodegenerative diseases. Research in her laboratory was/is funded by grants from Telethon Italia, the Italian Ministry of Health, the Italian Ministry of Education, University and Research, and the Autonomous Province of Trento.
Prof. Michela Alessandra Denti obtained her PhD in Biological Sciences in 1997 from the Scuola Normale Superiore di Pisa, Italy, working on the in vivo application and intracellular localisation of a hammerhead ribozyme and its association with RNA-binding proteins. She then went on to do postdoctoral research in Greece and at the University of Rome, Italy, studying the interaction of plant RNA pathogens with RNA-binding proteins and their involvement in post-transcriptional gene signalling and developing an antisense-U1snRNA mediated exon skipping therapy for Duchenne muscular dystrophy, respectively.