A Sensitive Alternative for MicroRNA In Situ Hybridizations Using Probes of 2 '-O-Methyl RNA plus LNA

作者：Soe Martin J; Moller Trine; Dufva Martin; Holmstrom Kim^*

来源：Journal of Histochemistry and Cytochemistry, 2011, 59(7): 661-672.

DOI：10.1369/0022155411409411

• in situ hybridization
• microRNA
• locked nucleic acids
• 2 '-O-methyl RNA
• yeast RNA
• urea
• formalin-fixed paraffin-embedded
• microRNA-138
• microRNA-205

The use of short, high-affinity probes consisting of a combination of DNA and locked nucleic acid (LNA) has enabled the specific detection of microRNAs (miRNAs) by in situ hybridization (ISH). However, detection of low-copy number miRNAs is still not always possible. Here the authors show that probes consisting of 2'-O-methyl RNAs (2OMe) and LNA at every third base (2:1 ratio), under optimized hybridization conditions, excluding yeast RNA from the hybridization buffer, can provide superior performance in detection of miRNA targets in terms of sensitivity and signal-to-noise ratio compared to DNA + LNA probes. Furthermore, they show that hybridizations can be performed in buffers of 4M urea instead of 50% formamide, thereby yielding an equally specific but nontoxic assay. The use of 2OMe + LNA-based probes and the optimized ISH assay enable simple and fast detection of low-copy number miRNA targets, such as miR-130a in mouse brain. (J Histochem Cytochem 59:661-672, 2011)

• 出版日期2011-7