miRNA Luciferase Reporter Assay

Services

CBpromise   

Our promise to you:
Guaranteed product quality, expert customer support.

24x7 CUSTOMER SERVICE
CONTACT US TO ORDER

miRNA Luciferase Reporter Assay

IntegrateRNA offers a high sensitive miRNA luciferase reporter assay service to help you quantitatively evaluate miRNA regulation activity. Simply sending us the information of your miRNA, then professional scientists will offer comprehensive solution, including miRNA target prediction, vector construction, cell transfection, luciferase activity detection and report delivery. We guarantee to provide our customers with high-quality service from experimental design to data analysis with faster turnaround time and lower prices.

MicroRNAs (miRNAs) regulate gene expression by targeting mRNA at specific sites in the 3 ́UTR, thereby inducing cleavage of the targets or inhibiting translation. The interaction of the miRNA with the target is mediated by an RNA-induced silencing complex (RISC), and depends on the sequence complementarity and the total number of binding sites in a given 3 ́UTR. Thus, to understand the roles of miRNA in complex biological processes, it is important to experimentally assess the functional relevance of the predicted miRNA targeting sites.

Cell assays based on the luciferase reporter gene can be adapted for testing the effect of miRNA-mediated, post-transcriptional regulation on target genes. As shown (Figure 1), the experimental procedure first consists in cloning both the wild-type and mutant forms of the 3'UTR of the miRNA predicted mRNA target downstream of a firefly luciferase reporter. Next, each construct is co-transfected together with the miRNA into suitable cell line, and the reporter expression is monitored. Changes in luciferase levels will indicate whether or not a miRNA can bind to the UTR and regulate its expression.

Figure 1. Principle of miRNA luciferase reporter assayFigure 1. Principle of miRNA luciferase reporter assay

Workflow of Luciferase Reporter Assay Service

  • Cloning the wild-type and mutant miRNA targeting site into luciferase reporter vector
  • Co-transfection of plasmids and miRNA mimic into suitable cell line
  • Measuring the activity of luciferase
  • Data analysis and report delivery

Applications

  • Monitoring miRNA-mediated regulation of cellular target genes
  • miRNA binding site analysis
  • Transcript stability study
  • 3'-UTR analysis

IntegrateRNA is a global leading biotechnology company specializing in RNA research. With skilled and experienced genomics experts, we are able to provide professional and efficient miRNA luciferase reporter assay to validate the predicted miRNA targets and detailed binding sites. If you have any questions, don't hesitate to contact us.

Reference:

  1. Yi Jin. et al. Evaluating the MicroRNA Targeting Sites by Luciferase Reporter Gene Assay. Methods in Molecular Biology, 2013, 936:117-127.
For research use only. Not intended for any clinical use.
Verification code