MicroRNAs (miRNAs) are a prevalent class of small single-stranded non-coding RNAs (19-30 nts long). They serve widespread functions as regulatory molecules in post-transcriptional gene silencing.
Since the discovery of miRNAs in 1993, the number of miRNAs in the Sanger miR database has rapidly increased. Approximately 4400 precursor miRNAs (based on miRBase) have been found to date in virtually all species—animals, plants, and viruses.
This diverse yet fundamentally conserved group of small RNAs may rival classical transcription factors in their role and regulatory circuitry found in cells. Much recent human cancer research has been intensely focused on studying and understanding miRNA expression.
Key Topics
| Components of the Agilent miRNA microarray probe design |
An unmodified microarray probe (black) is a synthesized sequence that hybridizes to the target miRNA (red). Probes are anchored to the glass slide surface by a stilt (brown).
- Inclusion of a G residue (black) to the 5' end of the hybridization sequence complements the 3' end C residue (yellow) introduced in labeling. This additional G-C pair in the probe-target interaction region stabilizes targeted miRNAs relative to homologous RNAs. Additionally, all probes contain a 5' hairpin (blue), abutting the probe-target region, to increase target and size miRNA specificity.
- Destabilization of probes that are too stable. For probes requiring it, reduction of probe-target base-pairing is achieved through sequential elimination of base pairing from the 5' end of the miRNA.
| Scientific technology behind Agilent's miRNA platform | The research of Wang et al. from the journal RNA.
The paper highlights key technological advances required for the development of a microarray-based method for profiling miRNAs. The scientific breakthroughs detailed in this paper provide an important foundation for the development of the Agilent platform.
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