GFP Application Areas
GFP is a versatile tool with a broad application base. Application areas include:
Research
Since its cloning in 1992 by Prasher and colleagues, GFP has been extensively and creatively utilized for research to elucidate the molecular mechanisms of cell biology.
D.C. Prasher et al., Primary structure of the Aequorea victoria green fluorescent protein. Gene 1992, 111:229-233.
Pharmaceutical screens
As demand shifts away from in vitro assays toward assays in living cells, the use of GFP reporters is becoming increasingly attractive for pharmaceutical screening, particularly in the areas of lead optimization and target validation. Pharmaceutical screens involving GFP generally monitor either protein behavior or gene activation in response to treatment with test compounds
Live-cell screening using the IN Cell Analyzer. GFP fusion protein to report transcription factor translocation.
Transgenic organisms
Multicellular organisms can be genetically engineered to express GFP, usually by introducing the DNA into germ line cells. Within a transgenic organism, the gene encoding GFP can be fused in-frame with the gene for a specific protein in order to follow the behavior of that protein in vivo. Alternatively, the gene encoding GFP can be placed under the control of a specific response element in order to study regulation of gene activation in vivo. Application areas for transgenic organisms include:
- Gene therapy
- Functional biology (genomics, proteomics)
- Cell engineering
- Therapeutic animal models
- Therapeutic protein production
 | GFP reporting activation of 'crystallin' genes in the lens of the frog eye (Xenopus). Image from Enrique Amaya's website. |
Functional biology
The human genome sequencing effort has given rise to a wealth of genes encoding proteins of unknown function. GFP technology can be exploited to report directly or indirectly on the behavior of target proteins or DNA elements.
Use of triGFP to localize expression of a cDNA of unknown function in live cells. U2OS cells were transiently transfected with (A) triGFP and (B) triGFP-cDNA fusion and imaged by epi-fluorescence microscopy.
Localization of a triGFP-cDNA fusion protein in U2OS cells by confocal microscopy with MitoTracker® red mitochondrial marker (Molecular Probes).
For a general review of GFP applications in cell biology and biotechnology, see Misteli T. and Spector D.L. Applications of the green fluorescent protein in cell biology and biotechnology. Nat. Biotechnol. 1997, 15(10):961-964. |
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