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We have previously used a defective herpes simplex virus vector to express a foreign gene in the adult rat brain. One application of this technology would be the in vivo analysis of promoter function in brain after de novo transfer, which would allow the rapid generation of vectors with localized application in a broad range of mammalian species while avoiding influences of other nearby promoters. A 2.7-kb fragment of the rat preproenkephalin promoter was placed upstream of the bacterial lacZ gene in our herpes simplex virus amplicon. A restricted pattern of lacZ expression was observed in vivo, which follows previously observed patterns of endogenous preproenkephalin expression. These results, from the direct gene transfer into an adult animal brain for in vivo promoter analysis, demonstrate that sequence information that influences restricted expression of preproenkephalin is located within 2.7 kb upstream of transcriptional initiation. lacZ expression was also observed in rat brain for 2 months after direct transfer, and PCR analysis confirmed the continued presence of amplicon DNA in lacZ-positive sections. Restricted and long-term expression observed with an endogenous promoter has important implications for gene therapy using viral vectors.

proceedings of the national academy of sciences of the united states of america

Preproenkephalin promoter yields region-specific and long-term expression in adult brain after direct in vivo gene transfer via a defective herpes simplex viral vector.