IntegrateRNA provides a specialized luciferase reporter assay service aimed at validating circRNA targets and uncovering detailed binding sites. This service leverages advanced molecular biology techniques and experienced professionals to deliver innovative solutions with dependable quality at competitive prices.
The Importance of circRNAs
Circular RNAs (circRNAs) are unique single-stranded transcripts characterized by their covalently closed structure, produced from precursor mRNA through back splicing in eukaryotes. The emergence of high-throughput RNA sequencing and cutting-edge bioinformatics tools has facilitated the identification of thousands of circRNAs across various species. Recent studies demonstrate that circRNAs can interact with RNA and proteins, acting as regulators of transcription or splicing. Notably, their roles as miRNA sponges have heightened the need for experimental validation of their interactions.
Mechanism of the Luciferase Reporter Assay
The luciferase reporter assay is a widely accepted method for investigating gene expression at the transcriptional level. This assay can also assess the regulation of miRNAs specific to circRNAs. The process typically involves the construction of a luciferase vector (e.g., psiCHECK2) where the target circRNA sequence is inserted into the 3’ UTR of a promoter-driven luciferase gene. Cells are then co-transfected with this luciferase reporter vector and miRNA mimics, enabling researchers to analyze luciferase activity in comparison to negative controls.
When miRNA binds to a specific circRNA, luciferase gene expression is negatively regulated. If the sequence of the circRNA binding site undergoes mutations that disrupt its sponge function, luciferase expression remains unaffected by miRNA mimics, providing critical insights into interaction dynamics.
Schematic diagram of circRNA luciferase reporter assay.
Workflow Overview
The circRNA luciferase reporter assay follows several key steps:
- Design and construct the luciferase vector containing the circRNA sequence.
- Co-transfect cells with the luciferase reporter vector and miRNA mimics.
- Measure luciferase activity to assess the interaction between miRNAs and circRNAs.
- Compare results with negative controls and analyze data for binding site validation.
Advantages of the Service
IntegrateRNA’s circRNA luciferase reporter assay service offers several significant benefits:
- Nearly instantaneous measurements.
- Exceptional sensitivity that enhances detection capabilities.
- A wide dynamic range suitable for various experimental conditions.
- Compatibility with most cell lines.
- A one-stop solution coupled with a quick turnaround time.
As a leader in biotechnology with a focus on RNA biology, IntegrateRNA incorporates expertise from genomics specialists to deliver comprehensive services. By submitting the sequence information for a specific circRNA and/or predicted miRNAs, clients can expect reliable and satisfactory experimental results efficiently.
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