A Novel Ligase-Based System to Efficiently Label MicroRNAs

The Rockefeller University
posted on 01/26/2010

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MicroRNAs are small RNA molecules (21 - 23 nucleotides in length) that act as regulators of gene expression in plants and animals, including humans. There is a need to characterize the expression patterns of these miRNAs in specific tissues and cell types in both healthy and disease states. One of the approaches for detecting miRNAs is based on microarray hybridization, which requires fluorescent labeling of the miRNA sample. Some of the current labeling methods use RNA ligases and fluorescently labeled donor nucleotides, but there are undesirable side reactions that result in a sequence dependent formation of circular miRNA species thus leading to a labeled pool that does not correctly represent the composition of the biological sample. Using a modified RNA ligase molecule our investigators have developed a novel system and corresponding protocol to minimize the side reactions during the labeling reaction. This allows direct labeling of natural 5

The detection and visualization of labeled miRNAs is useful for life sciences research and diagnostic applications.

The novel components in this system increase the yield of the desired linear species of labeled miRNAs and give a labeled pool that is a better representation of the composition of the biological sample, thus allowing a precise quantitative evaluation of the microarrays. This labeling technique also allows the incorporation of multiple labels in that may increase detection sensitivity.