Use of enhanced green fluorescent protein to optimize and quantitate infection of target cells with recombinant retroviruses.
Recombinant retroviral vectors are useful tools for gene transfer in both gene therapy and research applications. An enhanced form of green fluorescent protein has been incorporated into recombinant retroviruses as a marker to follow infected cells. In this paper, we extended the use of the fluorescent reporter to quantify protein expression using such analytical tools as fluorescent microscopy, flow cytometry and fluorescent plate reader analysis. These tools enabled us to rapidly assess the titer of recombinant retrovirus harvested from packaging cells and to optimize parameters for infection of different cell lines.
Cashion
L M
LM
Department of Biological Research, Berlex Biosciences, Richmond, CA 94804-0099, USA.
Bare
L A
LA
Harvey
S
S
Trinh
Q
Q
Zhu
Y
Y
Devlin
J J
JJ
D016475:3T3 Cells; D000818:Animals; D001709:Biotechnology; D002460:Cell Line; D005453:Fluorescence; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics*; D051379:Mice; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics*
D005819:Genetic Markers; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins