Rapid identification of viable retrovirus-transduced cells using the green fluorescent protein as a marker

Abstract
Various methods for determining the expression of the β-galactosidase (β-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 gener-ally superior to β-gal in terms of sensitivity, speed and noninvasiveness 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.

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