circRNA Luciferase Reporter Assay

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circRNA Luciferase Reporter Assay

IntegrateRNA provides you with luciferase reporter assay service for the circRNA targets validation and detailed binding sites discovery. Base on advanced molecular biology experiment platform and professional scientists in molecular biology experiment, we promise an innovative solution with reliable quality and competitive price for global customers. The ultimate goal of our service is to facilitate your basic research and drug discovery.

Circular RNAs (circRNAs) are covalently closed and single-stranded transcripts that are produced from precursor mRNA back splicing in eukaryotes. The improvements in high-throughput RNA sequencing and novel bioinformatics tools have led to the identification of thousands of circRNAs in various organisms. Recent findings have indicated that circRNAs may interact with RNA or proteins and serve as transcription or splicing regulators. Particularly, circRNAs have been found to interact with microRNA (miRNA) and function as miRNA sponges. Therefore, it is important to experimentally validate the predicted miRNA and detail binding sites for circRNA mechanism research.

The luciferase reporter assay is commonly used as a tool to study gene expression at the transcriptional level. It also can be utilized to investigate the regulation miRNA complementary to specific circRNA. As shown (Figure 1), the luciferase vector (such as psiCHECK2) is constructed by inserting the specific circRNA sequence in the 3' UTR into the promoter-driven luciferase reported gene. Cells are co-transfected with the luciferase reporter vector and the miRNA mimics in order to establish the characteristic of circRNA as miRNA sponges by comparing the luciferase activity with the negative controls. Similarly, mutant circRNA sequence could be cloned into luciferase vector to verify the detail binding sites interacted with miRNA.

Schematic diagram of circRNA luciferase reporter assay.Figure 1. Schematic diagram of circRNA luciferase reporter assay.

  • If miRNA binds to specific circRNA, the luciferase gene will be down-regulated. The luciferase expression level is inversely proportional to the intensity of miRNA-circRNA interaction.
  • If the circRNA binding site sequence with sponge function is mutated, the luciferase expression level will not be affected by the miRNA mimics.

Workflow of circRNA luciferase reporter assay

circRNA Luciferase Reporter Assay

Advantage

  • Measurements that are almost instantaneous
  • Exceptional sensitivity
  • A wide dynamic range
  • Suitable for most cell lines
  • One-stop solution
  • Short turnaround time

As a leading global biotechnology company specializing in RNA biology, IntegrateRNA recently introduced dual-luciferase reporter assay for circRNA research. With skilled and experienced genomics experts, we are able to provide professional and efficient circRNA luciferase reporter assay to investigate the targeting miRNAs and the detail binding sites for a specific circRNA. Simply providing the sequence information of specific circRNA and/or predicted miRNA, a satisfactory and reliable experimental result will be delivered to you.

For research use only. Not intended for any clinical use.
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