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What is a Reporter Gene?
Reporter genes are indispensable tools in molecular biology and genetics, allowing for detailed studies of gene regulation. Used in a wide range of scientific applications like drug discovery and environmental toxicology, reporter genes inform researchers of changes in the activity of their receptors of interest. In this article, we will explore reporter genes and their use in bioassays!
Reporter Genes and Reporter Cells
Types of Reporter Genes
A reporter gene encodes a protein that provides an easily measured output signal. Most commonly, the expressed product is a fluorescent protein, or an enzyme that catalyzes a colorimetric, luminescent, or fluorescent signal. Common examples of reporter genes include B-galactosidase, luciferase, and green fluorescent protein (GFP).1
How Reporter Gene Assays Work
Reporter genes are functionally linked to upstream genetic response elements that have been engineered to an entire native promoter region that is specifically activated by the receptor or signal transduction pathway of interest. Frequently, a vector expressing the receptor of interest is co-introduced into cells along with the promoter-reporter gene vector. The resulting cells, called reporter cells, are then used to screen test samples, compounds, or drugs to identify those with bioactivity as either agonists or antagonists of the target receptor. In this way, reporter genes provide a sensitive, easily quantifiable, surrogate measure of test compound-induced changes in receptor activity.
Why Reporter Gene Assays are Used
Reporter gene assays are crucial tools since their relevance spans various applications, providing quantifiable and often highly sensitive, nondestructive readouts of biological activity. Reporter gene assays are ideal for high-throughput screening (HTS) of test substances, allowing for the rapid testing of thousands of samples. HTS applications typically use automated systems that can process hundreds of assay plates and deliver robust quantitative measures of target receptor activities.
Reporter Gene Assays in Drug Discovery
In drug discovery, reporter gene assays are employed to screen large libraries of compounds for potential therapeutic effects. For instance, a reporter gene under the control of a cancer-specific promoter may be used to identify compounds that inhibit cancer cell proliferation.
Reporter Gene Assays in Toxicology
Reporter genes can also help assess the toxicity of chemicals and environmental pollutants. Cells engineered with reporters linked to stress- or DNA damage-response elements can reveal toxicity of a substance by measuring changes in reporter activity. This is useful for environmental monitoring and safety testing of new compounds.
INDIGO’s Luciferase Reporter Gene Assays
INDIGO Biosciences develops receptor-specific reporter assays which utilize the bioluminescent enzyme firefly luciferase as the reporter gene. INDIGO’s luciferase reporter cells incorporate the cDNA encoding beetle luciferase, a 62 kD protein originating from the North American firefly (Photinus pyralis). INDIGO’s reporter cell systems are engineered to provide optimal assay sensitivity and dynamic range when quantifying receptor activity.
INDIGO’s assays incorporate a detection reagent specially formulated to provide stable light emission for 90+ minutes after initiating the luciferase reaction, thus allowing assay plates to be processed in batch. By doing so, the signal output from all sample wells, from one plate to the next, may be directly compared within an experimental set.
If your aim is to perform cell-based target validation, pathway analysis, test compound/drug screening, or functional monitoring of environmental test samples, INDIGO’s portfolio of optimized, all-inclusive firefly luciferase reporter assay kits and assay services can help.
Works Cited
- Naylor, L. H. (1999b). Reporter gene technology: The future looks bright. Biochemical Pharmacology, 58(5), 749–757. https://doi.org/10.1016/s0006-2952(99)00096-9