Gfp Reporter Assay
Gfp Reporter Assay - Related Videos
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.
09:22
Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
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Cited by 11 •
2013
This protocol describes an experimental procedure for the rapid construction of artificial transcription factors (ATFs) with cognate GFP reporters and quantification of the ATFs ability to stimulate GFP expression via flow cytometry.
06:19
In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy
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Cited by 5 •
2017
This protocol describes how high-resolution imaging techniques such as spectral domain optical coherence tomography and scanning laser ophthalmoscopy can be utilized in small rodents, using an ophthalmic imaging platform system, to obtain information on retinal thickness and microglial cell distribution, respectively.
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.
10:40
Characterization of Adipocyte-Derived Extracellular Vesicle Secretion Using a CD63-GFP Reporter Mouse Model In Vivo and In Vitro
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2025
The protocol uses an adipocyte-specific CD63-GFP reporter mouse line to visualize and quantify the secretion of adipocyte-derived extracellular vesicles (EVs) and demonstrate their uptake by progenitor cells, revealing a paracrine pathway within adipose tissue.
FAQs
The promoter or regulatory element determines which gene-regulatory activity the assay reveals. When that control sequence becomes active, it drives production of GFP in the corresponding cells or developmental context. Choosing different regulatory elements can therefore expose distinct activation patterns during embryogenesis, tissue formation, or cell differentiation.
The timing of fluorescence provides a temporal record of regulatory activity. By following signal appearance during development, researchers can determine when a selected gene-control element becomes active and relate that timing to tissue formation or cell differentiation. This time-resolved view helps connect gene regulation with changing developmental patterns.
Depending on how the reporter is designed, GFP fluorescence can provide information about gene expression, protein localization, or cellular activity. Signal distribution may show where a regulatory program is active, while localization patterns can indicate where a protein is found. These readouts help connect molecular events with cell behavior and tissue organization.
Researchers select a promoter or regulatory element, place the GFP coding sequence under its control, and examine fluorescence in living specimens. Blue or ultraviolet excitation reveals the emitted green signal. Repeated imaging across developmental stages then captures when and where the reporter becomes active, producing a spatial and temporal expression pattern.
Live imaging allows developmental patterns to be followed as they emerge rather than assessed only at a single time point. Researchers can observe reporter activity in living embryos and relate changing fluorescence to tissue formation, cell differentiation, and evolving cellular organization. This makes the assay useful for connecting gene regulation with developmental progression.
In developmental biology, the assay can identify when and where selected regulatory programs operate during embryogenesis, tissue formation, and cell differentiation. Fluorescence patterns can also be compared with cell behavior and tissue organization over time. These observations help researchers examine how gene regulation is associated with the construction and specialization of developing tissues.