New sensitive and specific method for microRNA analysis

Robert Sjöback1, Lukáš Valihrach2, Mikael Kubista1,2
1TATAA Biocenter AB, Sweden; 
2Institute of Biotechnology, CAS, Czech Republic

MicroRNAs (miRNAs) are small non-coding RNAs that function as important biological regulators in viruses, plants, and animals (1). The single-stranded miRNAs regulate gene expression at the post-transcription level and the dysregulation of miRNAs is associated with various human diseases (2-4). Recent reports show microRNAs are abundant not only in tissues but also in body fluids and they show great potential as minimum-invasive biomarkers in the diagnosis and prognosis of cancers and other diseases (5, 6). However, there are two main challenges when analyzing miRNAs by molecular methods: 1) miRNAs are short, usually not more than 22 nucleotides, which is the length of conventional PCR primers; 2) closely related miRNAs may differ in only a single nucleotide position. Current methods approach these challenges by extending the length of the miRNA using either miRNA specific RT primers or by non-specific RT primers, which compromises specificity and sensitivity (7). Here we present a novel method for the detection and quantification of short nucleic acids that has higher sensitivity than current approaches with concomitant enhanced specificity.
1) He et al., Nat Rev Genetics 2004, 5: 522-31. 2) Esquela-Kerscher A & Slack FJ, Nat. Rev. Cancer 2006, 6: 259-69. 3) Michael et al., Mol. Cancer Res. 2003, 1: 882-91. 4) Dimmeler S & Nicotera P, EMBO Mol. Med. 2013, 5: 180-90. 5) Brase JC et al., Mol. Cancer 2010, 26: 306. 6) Chen et al., Cell Res. 2008, 18: 997-1006. 7) Mestdagh P et al., Nature Methods 2014, 11:809-815.

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