Abstract
This chapter discusses the design and construction of replication-defective herpes simplex virus type 1 (HSV-1) vectors for a variety of purposes. The chapter also discusses advantages and disadvantages for using HSV-1 as a vector. Recombinant HSV-1 vectors do not contain a mixed genetic population and are therefore uniform in their effects on infected cells. HSV-I has a wide target cell range and can infect nondividing cells. The HSV-1 genome can stably persist in neurons during a latent infection and produce the latency-associated transcript (LAT) RNAs. The virus can accept fairly large inserts of foreign DNA (15 kb or more), depending on the amount of nonessential viral DNA that has been deleted. It is also possible to insert multiple foreign DNA sequences into the HSV-1 genome. Three inherent disadvantages of replication-defective HSV-1 vectors are: (1) the large size of the HSV-1 genome requires more complex procedures for its manipulation than simple cloning; (2) although HSV-I can efficiently infect dividing cells, it makes little sense to use a replication-defective HSV-1 vector to infect such cells for long-term experiments, as the introduced DNA cannot be transmitted to all the daughter cells except in rare cases following recombination with the cellular genome; (3) HSV-I vectors may have the potential to cause reactivation of latent virus in vivo.
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