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Single HSV-amplicon Vector Mediates Drug-induced Gene Expression Via Dimerizer System.

S. Wang, Jeremy Petravicz, X. Breakefield
Published 2003 · Biology, Medicine

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A variety of viral vectors have been used to deliver genes into various tissues. Most have typically relied on either viral or cell-specific mammalian promoters to express transgenes. More recently, regulated promoter systems have been developed to fine-tune gene expression. Due to limited transgene capacity in most viral vectors, regulatory elements are typically subcloned into two separate vectors, which must be delivered simultaneously to a target cell. Here, we have cloned all the components of the rapamycin-based "dimerizer" system into the pantropic HSV-amplicon vector and used it to deliver and regulate red fluorescent protein (RFP) expression in cultured cells in a drug-dose-dependent manner. 293T/17 cells infected at an m.o.i. of 1 transducing unit/cell and induced with 20 nM rapamycin resulted in a 25-fold increase in RFP mRNA levels after 24 h as assessed by quantitative RT-PCR. However, due to a reduced ability to detect RFP optically, only a 5-fold induction in the number of RFP-expressing cells was noted by FACS analysis 48 h after infection. Further, there was at least 100-fold variation in the levels of RFP in individual, infected cells in the induced state. Gene induction in several neuronal models, including primary cell culture and organotypic cultures, as well as in rodent brain, was observed.
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