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Detecting miRNA & siRNA

Sensitive Solution Hybridization Assay For Small RNAs


In the past few years, interest in the identification, detection, and use of small RNA molecules has exploded. This interest has primarily stemmed from two interrelated lines of research. In one, small double-stranded RNAs (dsRNAs) called small interfering RNAs (siRNAs) have been used to silence the expression of specific genes at the post transcriptional level by a pathway known as RNA interference (RNAi). In the other, numerous small regulatory RNA molecules, referred to as microRNAs (miRNAs), have been shown to regulate target gene expression in various organisms. It is now becoming apparent that siRNAs and miRNAs are related molecules, sharing common processing pathways and maybe even functional mechanisms.

Currently, most miRNA researchers are analyzing miRNA expression patterns by Northern blot, a technique that is relatively insensitive and labor intensive. A few researchers performing gene silencing experiments also use this technique to analyze siRNA levels after RNAi induction, although the majority of researchers performing gene silencing experiments do not monitor siRNA levels at all. This may be due in large part to the inherent difficulties in detecting small RNAs with standard techniques. Here we discuss an optimized technique for the detection and quantitation of small RNAs. This technique allows easier monitoring of miRNA expression and siRNA levels in
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