Dual Reporter Assay
Dual Reporter Assay - Related Videos
12:55
High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay
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Cited by 45 •
2014
We present a rapid and inexpensive screening method for identifying transcriptional regulators using high-throughput robotic transfections and a homemade dual-glow luciferase assay. This protocol rapidly generates direct side-by-side functional data for thousands of genes and is easily modifiable to target any gene of interest.
13:48
Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay
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Cited by 6 •
2008
Here, we describe procedures for studying changes in phagocytosis-induced gene expression with a luciferase-based reporter gene approach using the Dual-GloTM Luciferase Assay System from Promega.
04:07
A Luciferase Reporter Assay to Study Translation Regulation in Poxvirus-Infected Cells
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2025
This video demonstrates an assay to study the translation regulation in poxvirus-infected cells. Uninfected and virus-infected cells were co-transfected with mRNAs encoding for firefly luciferase (Fluc) and Renilla luciferase (Rluc), with the Fluc mRNA containing a 5' Poly(A) leader sequence. Upon adding bioluminescent substrates on lysed cells, measure the signal from both the luciferases to identify the translational advantage of the Fluc mRNA in virus-infected cells, conferred by the leader...
02:01
Assessing Pseudovirus Infection Using Luciferase Reporter Assay
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2026
Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021).This video demonstrates a luciferase-based assay to quantify pseudovirus infection in epithelial cells. The pseudovirus encodes the luciferase gene, which is expressed in infected cells. After washing, and cells are incubated with lysis buffer at an ultra-low temperature, and then thawed to induce lysis.
11:03
Massively Parallel Reporter Assays in Cultured Mammalian Cells
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Cited by 63 •
2014
The genetic reporter assay is a well-established and powerful tool for dissecting the relationship between DNA sequences and their gene regulatory activities. Coupling candidate regulatory elements to reporter genes that carry identifying sequence tags enables massive parallelization of these assays.
FAQs
The signal ratio compares activity from the reporter connected to the regulatory sequence or pathway with the activity of the internal-control reporter in the same sample. A change in this relative value indicates altered regulatory activity after experimental treatment, while reducing the influence of differences in transfection efficiency, cell number, or sample handling.
An internal control helps separate biological regulation from technical variation. If samples receive different amounts of transfected material, contain different cell numbers, or experience inconsistent handling, both reporter measurements provide a basis for normalization. This paired measurement improves reproducibility and makes comparisons between experimental conditions more meaningful.
Linking one reporter to a selected regulatory sequence allows its activity to serve as a readout of a specific promoter or signaling pathway. Comparing normalized values across conditions can show whether cytokines, microbial components, or other treatments alter that regulatory response. The internal-control signal helps preserve interpretability when sample-to-sample technical variation occurs.
A single reporter measurement can change because of biological regulation or because the sample differed technically. A Dual Reporter Assay adds a second measurement from the same sample, allowing the regulatory signal to be evaluated relative to an internal control. This comparison supports more reliable interpretation of pathway activation than an unnormalized signal alone.
The experiment links one reporter to the regulatory sequence or pathway of interest and includes a second reporter as an internal control in the same sample. After applying the selected experimental condition, both reporter activities are measured. The signal from the regulatory reporter is then compared with the control signal to obtain a normalized readout.
In immunology and infection studies, the regulatory reporter can monitor promoter activation, host inflammatory signaling, or pathogen-responsive pathways. Researchers can compare normalized responses after exposure to cytokines or microbial components. Because the assay accounts for technical variation, it helps determine whether observed changes more likely reflect altered host or pathogen-related regulation.
Candidate therapeutics can be evaluated by measuring their effects on a reporter linked to an inflammatory or pathogen-responsive regulatory pathway. The internal-control measurement provides normalization across samples, helping distinguish treatment-related changes in pathway activity from differences in transfection efficiency, cell number, or sample handling. The resulting relative signal supports comparison among experimental conditions.