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The loss of expression with d2egfp was more pronounced in terminally differentiated cells of the peripheral blood (>30 fold) than in progenitor cells (five to 10 fold), indicating a stronger transcription of the retroviral promoter in progenitor cells and a predominant role of protein inheritance over de novo synthesis of transgenic protein in mature blood cells. This analysis reveals an important and differentiation dependent contribution of protein half life to the expression of retroviral vectors in hematopoietic cells, establishes d2EGFP as a more accurate reporter for determination of vector transcription, and also suggests that preclinical data obtained under conditions of high transduction rates or with vectors expressing stable reporter proteins require careful interpretation. For two reasons, these issues require special consideration: first, preclinical models of retroviral (including lentiviral) gene transfer are often performed with a MOI that may not be desirable for a clinical application, and second, stable reporter proteins such as enhanced green fluorescent protein (EGFP) have become increasingly popular for preclinical evaluation of vector performance.1,2,3,4.
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