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Application Stories

What are microRNAs?

MicroRNAs constitute a recently discovered class of non-coding RNAs that play key roles in the regulation of gene expression. Acting at the post-transcriptional level, these fascinating molecules may fine-tune the expression of as much as 30% of all mammalian protein-encoding genes.

Mature microRNAs are short, single-stranded RNA molecules approximately 22 nucleotides in length. MicroRNAs are sometimes encoded by multiple loci, some of which are organized in tandemly co-transcribed clusters.

Transcription and processing of microRNA

MicroRNA genes are transcribed by RNA polymerase II as large primary transcripts (pri-microRNA) that are processed by a protein complex containing the RNase III enzyme Drosha, to form an approximately 70 nucleotide precursor microRNA (pre-microRNA). This precursor is subsequently transported to the cytoplasm where it is processed by a second RNase III enzyme, DICER, to form a mature microRNA of approximately 22 nucleotides (Figure 1). The mature microRNA is then incorporated into a ribonuclear particle to form the RNA-induced silencing complex, RISC, which mediates gene silencing.

Figure 1
MicroRNA biogenesis. (Click for a larger image)

MicroRNA and gene expression

MicroRNAs usually induce gene silencing by binding to target sites found within the 3’UTR of the targeted mRNA. This interaction prevents protein production by suppressing protein synthesis and/or by initiating mRNA degradation. Since most target sites on the mRNA have only partial base complementarity with their corresponding microRNA, individual microRNAs may target as many as 100 different mRNAs. Moreover, individual mRNAs may contain multiple binding sites for different microRNAs, resulting in a complex regulatory network.

The function of microRNAs

MicroRNAs have been shown to be involved in a wide range of biological processes such as cell cycle control, apoptosis and several developmental and physiological processes including stem cell differentiation, hematopoiesis, hypoxia, cardiac and skeletal muscle development, neurogenesis, insulin secretion, cholesterol metabolism, aging, immune responses and viral replication. In addition, highly tissue-specific expression and distinct temporal expression patterns during embryogenesis suggest that microRNAs play a key role in the differentiation and maintenance of tissue identity.

MicroRNA as disease biomarkers

In addition to their important roles in healthy individuals, microRNAs have also been implicated in a number of diseases including a broad range of cancers, heart disease and neurological diseases. Consequently, microRNAs are intensely studied as candidates for diagnostic and prognostic biomarkers and predictors of drug response.

MicroRNA research

MicroRNAs were first reported in mammalian systems in 2001. In the latest release of miRBase (v.15), more than 14000 microRNAs have been annotated, highlighting the rapid growth of this field of research. However, the functions of most of these microRNAs still remain to be discovered.

The challenges of studying microRNAs are two-fold. First, microRNAs are very short (~22 nt). This means that traditional DNA-based methods are not sensitive enough to detect these sequences with any reliability. Second, closely related microRNA family members differ by as little as one nucleotide, emphasizing the need for high specificity and the ability to discriminate between single nucleotide mismatches.

Exiqon’s microRNA tools

Exiqon has pioneered the development of microRNA research and diagnostics tools with leading-edge products and services based on the proprietary Locked Nucleic Acid (LNA™) technology. By incorporating LNA™ into our products, we have significantly increased the affinity and specificity of our probes, inhibitors and primers for their microRNA targets, thereby addressing both challenges described above.
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