MicroRNAs (miRNAs) are endogenous non-coding RNAs of about 19-25nt in length and are widely involved in post-transcriptional regulatory activities. Most miRNAs are highly sequence-conserved, expression-temporal, and tissue-specific. From embryonic development, to apoptosis, and even to tumor growth, miRNAs play an important role in a range of physiological and pathological processes. With its leading miRNA technology team and experimental platform, RiboBio provides a full range of miRNA services to accelerate the research process.
With RiboBio’s pmiR-RB-Report™ reporter gene detection system, researchers are able to construct the 3’UTR region of the miRNA target gene, and the regulatory effect of miRNA on target genes can be detected by simply co-transforming the miRNA mimic with the constructed target gene vector. pmiR-RB-Report™ is specifically designed to detect the binding activity of miRNAs to target genes. It contains the T7 promoter-driven reporter fluorescence (hRluc), whose downstream constructs the 3’UTR region of the target genes. The detection of the fluorescence can verify whether the miRNA has a regulatory effect on the target; and calibration fluorescence (hluc) is used to monitor the transfection efficiency and the target expression efficiency, which can be used as stable internal references.
Examples
Luciferase reporter gene assay data showed higher activity after treatment with miR-7 inhibitor and decreased activity after miR-7 mimic treatment. Yu Q, et al. J CrohnsColitis. 2016.
Transfection of miR-9 inhibitor into SH-SY5Y and SK-N-SH cells increased luciferase activity, whereas Mutant 3′-UTR did not. Zhang H, et al. Mol Cancer Ther. 2012.
Notes：
Since the construction difficulty and experimental difficulty are significantly increased as the length of the gene 3’UTR increases, it is recommended that the construction length shall be no more than 2.5 kb.
