Luciferase Expression
Luciferase Expression - Related Videos
08:09
High-throughput Titration of Luciferase-expressing Recombinant Viruses
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Cited by 14 •
2014
This article presents a high-throughput luciferase expression-based method of titrating various RNA and DNA viruses using automated and manual liquid handlers.
03:23
Measurement of the In Vivo Burden of Bacteria Expressing Luciferase in an Insect Larva
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2025
This video showcases the in vivo bacterial burden measurement using Bacillus Calmette-Guérin, BCG, lux-infected insect larvae. The process involves larval homogenization, treatment with a luciferase substrate, and bioluminescence measurement to determine relative light units that correlate with bacterial burden.
10:38
Monitoring Cell-autonomous Circadian Clock Rhythms of Gene Expression Using Luciferase Bioluminescence Reporters
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Cited by 78 •
2012
Circadian clocks function within individual cells, i.e., they are cell-autonomous. Here, we describe methods for generating cell-autonomous clock models using non-invasive, luciferase-based real-time bioluminescence technology. Reporter cells provide tractable, functional model systems for studying circadian biology.
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.
03:38
Measuring Mycobacterium smegmatis Mistranslation Rates Using a GFP/Luciferase Reporter System
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2025
Source: Chen, Y., et al., Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems. J. Vis. Exp. (2019)This video demonstrates a method to quantify mistranslation in Mycobacterium smegmatis using an inducible GFP and secreted mutant luciferase reporter system. It shows how luminescence resulting from mistranslation is measured and compared to GFP fluorescence to evaluate the impact of a test molecule on translational accuracy.
FAQs
At the reaction level, luciferase catalyzes the oxidation of its compatible luciferin substrate. This chemical reaction releases photons, creating the light signal detected in an assay. Because substrate compatibility is essential, the observed emission depends on whether the luciferase and luciferin pair can react effectively, allowing enzyme activity to be translated into a measurable biological readout.
Light intensity can reflect both the amount of luciferase produced and the activity of that enzyme under the assay conditions. A stronger signal may therefore indicate greater expression, greater catalytic activity, or contributions from both. Interpreting the result requires connecting the luminescence measurement to the biological question, such as altered gene regulation or cellular response.
A luciferase gene can function as a reporter linked to regulatory control of interest. Changes in its emitted light then provide a quantitative indication of how strongly that control system is active. This design allows researchers to monitor promoter activity and gene regulation, while also extending the readout to signaling pathways and broader changes in cellular behavior.
A typical measurement begins with cells or tissue samples that produce luciferase, followed by exposure to the enzyme’s compatible luciferin substrate. The resulting photon emission is then measured as an assay signal. Comparing light output across samples or conditions provides a way to evaluate changes in expression or activity associated with the biological process under study.
The approach is useful when researchers need a sensitive, quantitative indication of changing gene expression. Applications described for luciferase assays include examining molecular mechanisms, monitoring signaling pathways, assessing cellular responses to drugs, and studying infection. Measurements can be made in living cells or tissue samples, making the method relevant to both dynamic and sample-based experiments.
Researchers can use changes in emitted light to track how cells respond when a relevant biological condition changes, including exposure to a drug or involvement in infection. The assay converts those changes into quantitative measurements that can be compared between conditions. This helps connect molecular regulation with observable cellular behavior without relying only on qualitative inspection.