PMID: 9546076Apr 18, 1998Paper

Applications of green fluorescent protein as a marker of retroviral vectors

Somatic Cell and Molecular Genetics
Eugene S KandelI B Roninson

Abstract

The Green Fluorescent Protein (GFP) of Aequorea victoria is used as a vital fluorescent tag for the detection and isolation of genetically modified cells. Several modified variants of GFP were tested as marker genes in retroviral vectors containing different backbones and promoter combinations. Constructs allowing for reliable detection of GFP fluorescence and the expression of a cotransduced gene from a strong promoter were identified. Cells harboring such constructs are detectable by flow cytometry, fluorescence microscopy and multi-well fluorescence reading. GFP expression in transduced cells is stable both in vitro and in vivo, and long-term dynamics of GFP-positive fractions in a mixed population can be used to monitor the biological effects of a cotransduced gene. Selection of cells with the highest GFP fluorescence enriches for multiply infected cells. The use of different GFP variants allows one to monitor simultaneously two cell populations transduced with vectors carrying GFPs that differ in their fluorescence intensity or spectral properties and to identify doubly transduced cells. In addition, transcription of an inducible promoter positioned in the opposite orientation to GFP can be monitored by the inhibition of G...Continue Reading

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Citations

Feb 28, 2003·Analytical Biochemistry·Gerda EndemannDaria Mochly-Rosen
May 9, 2007·Oncogene·E V BroudeI B Roninson
Apr 27, 2005·Proceedings of the National Academy of Sciences of the United States of America·Eugene S KandelGeorge R Stark
May 12, 2010·Proceedings of the National Academy of Sciences of the United States of America·Zoya N DemidenkoMikhail V Blagosklonny
Oct 23, 2002·Molecular and Cellular Biology·Eugene S KandelNissim Hay
Apr 26, 2011·Journal of Bacteriology·Nikhilesh S ChandDeborah T Hung
Jul 13, 2002·Cytometry·Burt RichardsDavid H-F Teng
Apr 12, 2002·Brain Research. Developmental Brain Research·Christiana M Cooper-Kuhn, H Georg Kuhn

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