The individual gene expression blocking is miRNA general mechanism. In 2006 American scientists Andrew Fire and Craig Mello received the Nobel Prize in Physiology and Medicine for describing this phenomenon. Over the next years, our knowledge of the gene expression regulation system with the help of miRNA molecules has significantly extended. It is suggested that the complex system of these regulatory molecules interaction is the general mechanism for the dynamic regulation of the protein-secreting cell apparatus.
Neoplastic transformation occurs due to a composition change and, thus, due to the miRNA apparatus regulatory activity distortion. Moreover, this causal intercommunication can have different cause-effect relations.
It was noticed that the genome fragile sites – the sites of frequent aberrations and mutations – often encode entire miRNA molecules families. Mutations in these sites distort the composition of cellular miRNAs and lead to the development of various oncological diseases, including lung, colon and stomach cancer. This points to a possible etiological role of miRNA in cancer development. In addition, miRNAs in a normal cell regulate metabolism, proliferative activity, differentiation, and scheduled cell death (apoptosis). Disruption of these processes during neoplastic transformation can cause adaptive (secondary) changes in the intracellular miRNAs profile. The cancerogenic nature of some miRNAs has been confirmed by many studies. For example, miR-21, a miRNA molecule that inhibits the production of a number of tumor suppressor proteins by a cell, is involved in the development of many tumors. In modern molecular oncology, the concept of “onco-miR” has appeared—a miRNA with a distinctive cancerogenic effect, for example miR-155, a cluster of related molecules miR-106b / miR-17–92.
Neoplastic transformation of cells is accompanied by specific changes in the profile (composition) of intracellular miRNAs, which determines its potential diagnostic value. miRNAs isolation from biopsy specimen (fresh, fixed, stained) and analysis of the marker miRNA molecule profile is a promising diagnostic test. Moreover, the features of the miRNA profile can be associated with certain clinically significant characteristics of the tumor (for example, sensitivity to certain therapy), which can be used for personalization of treatment.
miRNA Assay Kits
A modern technological solution for basic research and development of new clinical diagnostic methods!
Assay without normalization
The possibility to quantitatively evaluate miRNA in RNA samples isolated from biological material appears after a calibration curve deriving. The calibration curve directly correlates amplification efficiency (threshold cycle, Ct) with analyte concentration (miRNA).
The concentration evaluation of miRNA under study might be carried out directly, without using the “normalization” procedure as to conditional «normalizers» (U6, U18, RNU44, RNU48, etc.), if the standardized RNA extraction protocol is used and Ct values in biological sample analysis fit into a linear dependency zone.
X‑direction: кoncentration of synthetic miR-371–3p molecules in the reaction mixture, 10^
RT-PCR is the main miRNA assay method in laboratory practice due to its reliability and economic efficiency.
The kit implements the original technology of reverse transcription two-way priming and PCR with two miRNA-specific primers (Androvic et al., 2017), that provides exceptional analysis accuracy.
Reliable results and higher assay sensitivity (compared to other technologies, Korobkina et al., 2018) are provided by RT primer structure optimization and amplification detection with a probe. This allows to work with low concentration RNA samples.
Inland manufactured optimized Enzyme Blends for RT and PCR.
Balanced mixtures of oligonucleotides for priming reactions and detecting the real-time amplification process.
A set of 8 calibrators—dilutions of synthetic “mimic” for making a calibration curve and miRNA assay in biological material.
Two phases of miRNA assay
Microarray technology, sequencing, including genome-wide RNA sequencing, and reverse transcription followed by polymerase chain reaction (RT-PCR) – the same methods that are traditionally used for the messenger RNA assay – are fundamentally applicable for the miRNA assay. Currently, RT-PCR is the most commonly used technique due to its reliability and relatively low cost.
Despite its widespread use, the practical use of the RT-PCR method is a sophisticated task.
- The miRNA assay results significantly depend on the quality of the RNA isolated from biological samples.
- The RT-PCR results can be distorted by the presence of molecules immature forms.
- The length of the mature miRNA molecule is comparable in size to the length of the primers, the pair of which must have “landing sites” along the detected molecule to initiate the polymerase chain reaction.
Reverse transcription (RT)
Ready to Use Reagents for Reverse Transcription
Polymerase chain reaction (PCR)
Ready to Use Reagents for PCR
miRNA RT-PCR Kits «AL-miR-RT-qPCR/hsa-*****»
Kits ALMIR available for order
Kits ALMIR available