Pushing miRNA quantification to the limits: high-throughput miRNA gene expression analysis using single-molecule detection
Authors: Puleo, Christopher M; Liu, Kelvin; Wang, Tza-Huei
Source: Nanomedicine, 1 June 2006, vol. 1, no. 1, pp. 123-127(5)
This study extended single-molecule detection (SMD) techniques to accurately quantitate microRNA (miRNA) gene expression in human tissue. In all, the expression of 45 different miRNAs was quantified from 16 different tissues and several mature miRNAs were found to be expressed at low levels in tissues where they were undetectable previously. This SMD assay showed the capability of quantitating miRNA expression from as little as 50 ng of total RNA. Furthermore, by incorporating locked nucleic acid (LNA)–DNA oligonucleotide probes, this method was shown to be highly specific and capable of discrimination between miRNA targets that differed by as little as a single nucleotide. Future extensions of SMD miRNA assays and integration with current microfluidic and nanotechnologies may prove essential in pushing miRNA profiling to the fundamental limits for discerning variation in the spatiotemporal regulation of miRNA expression across various tissue and cell types, as well as disease states, and establishing the clinical utility of miRNA expression in cancer diagnosis.
Document Type: Research Article
Affiliations: 1Johns Hopkins University, Biomedical Engineering Department and Whitaker Biomedical Engineering Institute, Baltimore, MD 21218, USA
Publication date: June 1, 2006