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  • Since their discovery a little over 20 years ago, miRNAs have emerged as an exciting and promising research and applied science target due to their involvement in many important cellular processes and their clear links to disease states. Bioline has developed a range of products to support miRNA research, including a range of expertly designed qPCR panels for analyzing important miRNAs in the areas of cancer, stem cells and biofluid analysis.

    miRNAs and Methodologies for Detection

    There is a diverse collection of miRNAs now discovered across a range of different organisms, from worms to mammals, flies and also plants. These small (16-25 nucleotide) non-coding RNAs have been found to participate in the regulation of important cellular functions including cellular development, differentiation, proliferation and apoptosis.

    Studies in the human genome have seen dysfunction in miRNA regulation linked to many different disease states and research continues in attempts to unravel the complex ways in which miRNAs interact with other genetic elements affecting gene expression on a number of different levels.

    The most studied mechanism of miRNA mediated gene regulation involves the mature miRNA and an RNA-induced silencing complex, this miRNA-RISC complex then binds to mRNA leading to degradation or translation inhibition. Each miRNA has the potential to target a number of different genes, plus research has also demonstrated ribonucleoprotein binding in a RISC-independent manner, as well as direct binding to DNA.1 Thus the regulation of gene expression by miRNAs is a complex and multi-level phenomenon.

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    qPCR Analysis of miRNAs

    There are a number of different ways to approach the study and profiling of miRNAs and real time PCR (qPCR) remains the most popular choice given its accessibility and potential for sensitive and specific results.

    It is not a straight forward practice, however, as qPCR analysis of miRNAs requires a number of challenges to be overcome including:

    • Short length of miRNAs
    • Target sequence present in pre-cursor forms as well as the mature miRNA
    • Closely related miRNAs can differ by as little as a single nucleotide
    • GC content across miRNAs is very heterogeneous
    • Difficulty in selectively isolating/purifying miRNAs from mixed RNA populations

    Bioline have developed a unique 3-primer strategy for their miRNA assays that clearly differentiate mature miRNAs as well as discriminate between closely related family members. Using SYBR® Green-based detection, the protocol is a streamlined and sensitive way to study individual miRNA, or larger scale miRNA panels (EPIK™ miRNA panel assays).

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    Genome-wide Analysis of miRNAs

    Deep sequencing using next generation sequencing (NGS) platforms is another popular method for profiling miRNA populations and, more importantly, identifying and discovering novel miRNAs. NGS technologies typically produce read lengths compatible with the length of mature miRNAs and the direct transcript count allows for determination of expression levels based on abundance.


    Trends and Applications in miRNA Research

    miRNA regulation (or dysregulation) has been studied in relation to a range of diseases including cardiovascular disease, psychiatric disorders, diabetes and cancers and as research into the full range of regulatory interaction continues, the list of diseases continues to grow.

    The connection between specific miRNAs and disease state is not surprising, given the clearly important role they play in most cellular processes and studies have now shown tissue-specific miRNA expression profiles that directly link to the physiological state of the originating cell. Moreover, comprehensive miRNA expression profiling has identified disease-specific expression patterns reflecting the lineage and differentiation status of various cancers. Thus miRNA-based research holds significant promise in the applications of prognostic and diagnostic oncology and many other therapeutic areas. Bioline offers a comprehensive panel of cancer-related miRNAs in their EPIK™ cancer miRNA qPCR panels to simplify and support research and application efforts within this field.

    miRNAs have been classified as oncogenes, with a demonstrated role in the negative regulation of tumor suppressor genes, as well as controlling cellular differentiation, proliferation and survival. Found in a range of different biological fluids including plasma, serum and saliva, miRNAs exhibit excellent stability thanks to their small size and presence within microvesicles, as well as other potential mechanisms that protect them from the degradation that often affects other circulating RNA. These characteristics in conjunction with the cell-specific information they represent, have led miRNAs to be a leading candidate in biomarker studies and Bioline offers a qPCR panel representing the most important miRNAs found in biofluids.

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    1. Iorio MV & Croce CM. MicroRNA dysregulation in cancer: diagnostics, monitoring and therapeutics. A comprehensive review. EMBO Mol Med 2012:4(3);143-159.

    Our complete miRNA research portfolio includes: