Egfp Reporter Expression
Egfp Reporter Expression - Related Videos
10:03
Development of Leishmania Species Strains with Constitutive Expression of eGFP
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2023
Here, we describe the methodology used for generating L. panamensis and L. donovani strains expressing the gene for eGFP as a stable integrated transgene using the pLEXSY system. Transfected parasites were cloned by limiting dilution, and clones with the highest fluorescence intensity in both species were selected for further use in drug screening assays.
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.
03:57
Preparation of Reporter-Expressing Cancer Cells for In Vivo Imaging
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2026
Source: Chen, H. et. al. Live Cell Imaging of the TGF-β/Smad3 Signaling Pathway In Vitro and In Vivo Using an Adenovirus Reporter System. J. Vis. Exp. (2018)This video demonstrates the preparation of luciferase-expressing breast cancer cells using recombinant adenoviruses for in vivo imaging.
10:48
Vaccinia Reporter Viruses for Quantifying Viral Function at All Stages of Gene Expression
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Cited by 5 •
2014
We describe the usage of a fluorescent reporter vaccinia virus that enables real-time measurement of viral infectivity and gene expression through the stage-specific expression of spectrally distinct reporter fluorophores. We detail a plate-based method for accurately identifying the stage at which virus replication is affected in response to small molecule inhibition.
10:30
Visualization of IL-22-expressing Lymphocytes Using Reporter Mice
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Cited by 2 •
2017
We describe here a transgenic reporter mouse model to visualize the IL-22-producing cells inside different mouse tissues. This method can be used to track the location of other cytokines or secretary proteins in the mouse.
FAQs
Promoter or regulatory element choice determines which genetic program drives EGFP production. When that control sequence is active, it directs transcription of the linked coding sequence, followed by translation into protein. Changing the regulatory element therefore changes the biological pattern represented by fluorescence, allowing researchers to relate signal location to specific gene-regulatory activity rather than treating green labeling as a nonspecific stain.
The chromophore is the light-responsive part of EGFP that fluoresces after excitation with blue light. It forms within the protein, so the observed signal depends on successful production of the reporter protein as well as suitable illumination. This mechanism gives researchers an optical readout that can be captured by fluorescence microscopy and connected to the location of active genetic programs.
Fluorescence microscopy shows where EGFP signal occurs, whereas quantitative image analysis makes that signal measurable across cells and tissue regions. This pairing helps researchers connect the distribution of reporter fluorescence with gene regulation and compare patterns associated with neural development, connectivity, or disease-related changes. The resulting analysis extends observation beyond a simple visual label.
A basic workflow includes selecting a promoter or regulatory element, linking it to the EGFP coding sequence, and examining the resulting fluorescence with microscopy. Researchers can apply this construct in cultured cells or living tissue, then use image analysis to assess where signal appears. The workflow connects genetic design, protein production, optical detection, and interpretation of neural or other biological patterns.
In neuroscience, EGFP labeling can identify neuronal populations, reveal cell morphology, and trace projections. These uses allow investigators to examine which cells are targeted by genetic manipulations and how labeled structures are arranged in neural tissue. Because the same signal can be examined in cultures or living tissue, the approach supports studies of neural development, connectivity, and changes associated with disease.
Spatial fluorescence patterns can indicate where a selected genetic program is active and which neuronal cells or structures carry the reporter. Image-based measurements can then relate those patterns to morphology, projections, connectivity, or developmental and disease-related changes. The signal does not stand alone as a biological conclusion; its meaning comes from the promoter or regulatory element chosen and the tissue context examined.