27 - Expression of Neurotrophic Genes from Herpes Simplex Virus Type 1 Vectors: Modifying Neuronal Phenotype

Abstract

Publisher Summary This chapter focuses on expression of neurotrophic genes from herpes simplex virus type 1 (HSV-1) vectors. To date, HSV-based vectors are one of only three means of transferring genes into postmitotic cells such as neurons. HSV-1 is ideally suited as a vector for gene transfer for many reasons. It can infect a variety of cell types in many different organisms. Its large size (150 kb) suggests that HSV-1 vectors can be produced to carry relatively large genes. It can persist indefinitely in a latent state in neurons without affecting their electrophysiological properties, and HSV-1 genes are transcribed by cellular RNA polymerase II, suggesting that cellular promoters placed in the vector could be transcribed as well. This chapter discusses methodologies used for one type of HSV-1 defective viral vector, the amplicon. This is a plasmid-based system of gene transfer that is effective for many cell types, including postmitotic neurons. The chapter presents methods for growing, titering, and applying defective HSV-1 vectors, both in vitro and in vivo . These viral vectors are defective, nonreplicating HSV-l-derived viruses. The usefulness of these vectors for transducing genes to modify neuronal function is illustrated by vectors carrying neurotrophic factor genes.