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Fluorescent Tracer
GFP has been used successfully to track the efficiency of gene delivery, both to cells in culture and to whole organisms. Typically, GFP is co-expressed with the gene of interest either bi-cistronically or on a separate delivery vector. In the latter case, the ratio of target gene to GFP may be adjusted to ensure that all cells expressing GFP have a high probability of expressing the gene of interest. GFP has also proved to be a useful biological tracer to monitor cell lineage and to improve techniques for the production of genetically modified organisms. Examples of the use of GFP as a fluorescent tracker include:
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Developing methods for gene therapy
A. Bartholomew et al., Baboon mesenchymal stem cells can be genetically modified to secrete human erythropoietin in vivo. Hum. Gene Ther. 2001, 12(12):1527-1541.
M. Goerner et al., Sustained multilineage gene persistence and expression in dogs transplanted with CD34(+) marrow cells transduced by RD114-pseudotype oncoretrovirus vectors. Blood 2001, 98(7):2065-2070.
Tracing cell lineage in the development of cell- and organ-transplant therapies
J. Priller et al., Neogenesis of cerebellar Purkinje neurons from gene-marked bone marrow cells in vivo. J. Cell Biology 2001, 155(5):733-738.
S. Wu et al., Migration, integration and differentiation of hippocampus-derived neurosphere cells after transplantation into injured rat spinal cord. Neurosci. Lett. 2001, 312(3):173-176.
Y. Hakamata et al,. Green fluorescent protein-transgenic rat: a tool for organ-transplantation research. Biochem. Biophys. Res. Commun. 2001, 286(4):779-785.
Developing improved somatic nuclear transfer techniques for animal cloning
S. Roh et al., In vitro development of green fluorescent protein (GFP) transgenic bovine embryos after nuclear transfer using different cell cycles and passages of fetal fibroblasts. Reprod. Fertil. Dev. 2000, 12(1-2):1-6.
Production of transgenic organisms
T. Takada et al., Selective production of transgenic mice using green fluorescent protein as a marker. Nature Biotechnology 1997, 15:458-61.
M. Ikawa et al,. A rapid and non-invasive selection of transgenic embryos before implantation using green fluorescent protein (GFP). FEBS Lett 1995, 375(1-2):125-8.
Following tumor metastasis and neoplastic disease
T. Chishima et al., Governing step of metastasis visualized in vitro. Proc.Natl. Acad. Sci. USA 1997, 94(21):11573-11576.
C.H. Contag et al., Use of reporter genes for optical measurements of neoplastic disease in vivo. Neoplasia 2000, 2(1-2):41-52.
R.M. Hoffman. Visualization of GFP-expressing tumors and metastasis in vivo. Biotechniques 2001. 30(5):1016-1026.
- Isolating cell populations expressing an untagged gene of interest
- Monitoring the efficiency of gene delivery in transiently transfected cell populations
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