Luciferase Gene Transcription
Luciferase Gene Transcription - Related Videos
08:55
In vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection
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Cited by 13 •
2012
This method describes high yield in vitro synthesis of both capped and uncapped mRNA from a linearized plasmid containing the Gaussia luciferase (GLuc) gene. The RNA is purified and a fraction of the uncapped RNA is enzymatically capped using the Vaccinia virus capping enzyme. In the final step, the mRNA is transfected into HeLa cells and cell culture supernatants are assayed for luciferase activity.
12:58
Transfecting RAW264.7 Cells with a Luciferase Reporter Gene
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Cited by 13 •
2015
Transfection into the macrophage cell line, RAW264.7, is difficult due to the cell’s natural response against foreign materials. We described here a gentle yet robust procedure for transfecting luciferase reporter genes into RAW264.7 cells.
09:44
High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
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2015
Here we describe a cell-based reporter gene assay as a valuable tool to screen chemical libraries for compounds modulating post-transcriptional control mechanisms exerted through 3’ UTR.
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.
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.
FAQs
When a regulatory sequence activates transcription, the luciferase gene produces messenger RNA, which directs production of the luciferase enzyme. After substrate is supplied, that enzyme generates light. The resulting luminescence provides an indirect readout of regulatory activity, allowing researchers to connect changes in transcription with measurable differences in signal rather than relying only on molecular detection of RNA.
The substrate provides the reactant required for luciferase to produce light. It does not represent the regulatory sequence itself; instead, it converts the enzyme produced after transcription into an observable luminescent output. Supplying substrate therefore links reporter expression to detection and makes transcriptional differences accessible through light measurement.
Researchers can place different promoters or regulatory sequences in comparable luciferase reporter designs and examine their luminescent outputs. Stronger transcriptional activation generally produces more reporter enzyme and a larger signal under the same measurement context. This comparison helps evaluate promoter strength and determine how regulatory elements influence gene expression.
Repeated luminescence measurements can show how reporter output changes as cellular regulation proceeds. This time-resolved perspective helps researchers examine responses to signaling activity, hormones, drugs, or environmental conditions rather than limiting analysis to one endpoint. Changes in signal over time can reveal when regulatory effects emerge and how long they persist.
A typical workflow connects a regulatory sequence to a luciferase reporter, allows transcriptional regulation to occur, supplies the luciferase substrate, and measures the emitted light. The signal is then interpreted as an indicator of reporter activity. This sequence of steps links regulatory control, enzyme production, chemical light generation, and quantitative detection in one assay.
These assays can evaluate promoter performance, regulatory sequence activity, signaling pathway effects, and cellular responses to hormones, drugs, or environmental conditions. They also help assess whether a genetic construct produces the intended regulatory behavior. In biological techniques, the approach is valuable because one luminescent readout can support comparisons across gene-regulation experiments.