Various methods for determining the expression of the beta-galactosidase (beta-gal) gene after retroviral transduction were compared as a means to assess retroviral titre. To allow better comparison, different retroviral vectors were constructed carrying two mutants of the green fluorescent protein and assessed as sensitive markers of retroviral gene transfer. It could be demonstrated that GFP is generally superior to beta-gal in terms of sensitivity, speed and non-invasiveness of assay, allowing easy direct FACS sorting of populations of transduced cells. This opens the possibility of enrichment by sorting of ex vivo transduced cells in gene therapy protocols.
Animals Cell Line Cell Separation Flow Cytometry Gene Transfer Techniques* Genetic Markers* Genetic Vectors* Green Fluorescent Proteins Humans Luminescent Measurements Luminescent Proteins*/genetics* Retroviridae* beta-Galactosidase/genetics*