Key result
Antibiotic resistance gene linkage constructs large HSV-1 vectors co-expressing multiple genes in rat brains.
A general strategy using genetic selections allows the construction of large HSV-1 plasmid vectors capable of co-expressing multiple genes in neurons.
Herpes simplex virus type 1 (HSV-1) plasmid vectors have a number of attractive features for gene transfer into neurons. In particular, the large size of the HSV-1 genome suggests that HSV-1 vectors might be designed to co-express multiple genes. Here, we report a general strategy for constructing large HSV-1 plasmid vectors that co-express multiple genes. Each transcription unit is linked to an antibiotic resistance gene, and genetic selections are used to assemble large vectors. Using this strategy, we constructed large (26 or 31 kb) HSV-1 vectors that contain two transcription units and two or three genes. These vectors were efficiently packaged into HSV-1 particles using a helper virus-free packaging system. The resulting vector stocks supported the expression of two or three genes in both cultured cells and the rat brain. Potential applications of HSV-1 vectors that co-express multiple genes are discussed.
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Wang et al. (2001) studied this question. Large HSV-1 plasmid vectors co-expressing multiple genes was evaluated on Expression of two or three genes in cultured cells and rat brain. A general strategy using antibiotic resistance gene linkage successfully constructed large HSV-1 plasmid vectors that co-expressed multiple genes in cultured cells and rat brain.
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