Construction and production of recombinant herpes simplex virus vectors.
Virus vectors have been employed as gene transfer vehicles for various pre-clinical and clinical gene therapy applications. Replication-competent herpes simplex virus (HSV) vectors that replicate specifically in actively dividing glial tumor cells have been used in Phase I-II human trials in patients with glioblastoma multiforme (GBM), a fatal form of brain cancer. Research during the last decade on the development of HSV vectors has resulted in the engineering of recombinant vectors that are totally replication defective, non-toxic, and capable of long-term transgene expression. This chapter describes methods for the construction of recombinant genomic HSV vectors based on the HSV-1 replication-defective vector backbones, steps in their purification, and their small-scale production for use in cell culture experiments as well as studies in animals.
Goins
William F
WF
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Krisky
David M
DM
Wechuck
James B
JB
Huang
Shaohua
S
Glorioso
Joseph C
JC
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004279:DNA, Viral / Q000235:genetics; D005822:Genetic Vectors / Q000096:biosynthesis*; D016679:Genome, Viral / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D014162:Transfection; D014771:Virion; D014779:Virus Replication
D004279:DNA, Viral