title: Construction and packaging of herpes simplex virus/adeno-associated virus (HSV/AAV) Hybrid amplicon vectors.
abstract: Herpes simplex virus type 1 (HSV-1)-based amplicon vectors conserve most properties of the parental virus: broad host range, the ability to transduce dividing and nondiving cells, and a large transgene capacity. This permits incorporation of genomic sequences as well as cDNA, large transcriptional regulatory sequences for cell-specific expression, multiple transgene cassettes, or genetic elements from other viruses. Hybrid vectors use elements from HSV-1 that allow replication and packaging of large-vector DNA into highly infectious particles, and elements from other viruses that confer genetic stability to vector DNA in the transduced cell. For example, adeno-associated virus (AAV) has the unique ability to integrate its genome into a specific site on human chromosome 19. The viral rep gene and the inverted terminal repeats (ITRs) that flank the AAV genome are sufficient for this process. However, AAV-based vectors have a very small transgene capacity and do not conventionally contain the rep gene to support site-specific genomic integration. HSV/AAV hybrid vectors contain both HSV-1 replication and packaging functions and the AAV rep gene and a transgene cassette flanked by the AAV ITRs. This combines the large transgene capacity of HSV-1 with the capability of site-specific genomic transgene integration and long-term transgene expression of AAV. This protocol describes the preparation of HSV/AAV hybrid vectors using a replication-competent/conditional, packaging-defective HSV-1 genome cloned as a bacterial artificial chromosome (BAC) to provide helper functions for vector replication and packaging. The advantages and limitations of such vectors compared to standard HSV-1 amplicon vectors are also discussed.
authors.forename: Okay
authors.initials: O
authors.lastname: Saydam
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Glauser
authors.forename: Cornel
authors.initials: C
authors.lastname: Fraefel
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D000229:Dependovirus / Q000235:genetics / Q000502:physiology*; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D058507:Host Specificity; D008967:Molecular Biology / Q000379:methods*; D011995:Recombination, Genetic; D014161:Transduction, Genetic; D014709:Vero Cells; D014773:Virology / Q000379:methods; D019065:Virus Assembly