Abstract
Amplicon vectors, or amplicons, are defective, helper-dependent, herpes simplex virus type 1 (HSV-1)-based vectors. The main interest of amplicons as gene transfer tools stems from the fact that the genomes of these vectors do not carry protein-encoding viral sequences. Consequently, they are completely safe for the host and nontoxic for the infected cells. Moreover, the complete absence of virus genes provides a genomic space authorizing a very large payload, enough to accommodate foreign DNA sequences up to almost 150-kbp, the size of the HSV-1 genome. This transgene capacity can be used to deliver complete gene loci, including introns and exons, as well as long regulatory sequences conferring tissue-specific expression or stable maintenance of the transgene in proliferating cells. For many years the development of these vectors and their application in gene transfer experiments was hindered by the presence of contaminating toxic helper virus particles in the vector stocks. In recent years, however, two different methodologies have been developed that allow generating amplicon stocks either completely free of helper particles or only faintly contaminated with fully defective helper particles. This chapter describes these two methodologies.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Spaete RR, Frenkel N (1982) The herpes simplex virus amplicon: a new eukaryotic defective-virus cloning-amplifying vector. Cell 30:295–304
Vlazny DA, Frenkel N (1981) Replication of herpes simplex virus DNA: localization of replication recognition signals within defective virus genomes. Proc Natl Acad Sci U S A 78:742–746
Spaete RR, Frenkel N (1985) The herpes simplex virus amplicon: analyses of cis-acting replication functions. Proc Natl Acad Sci U S A 82:694–698
Boehmer PE, Lehman IR (1997) Herpes simplex virus DNA replication. Annu Rev Biochem 66:347–384
Kwong AD, Frenkel N (1984) Herpes simplex virus amplicon: effect of size on replication of constructed defective genomes containing eukaryotic DNA sequences. J Virol 51:595–603
Fraefel C, Song S, Lim F, Lang P, Yu L, Wang Y, Wild P, Geller AI (1996) Helper virus-free transfer of herpes simplex virus type 1 plasmid vectors into neural cells. J Virol 70:7190–7197
Saeki Y, Fraefel C, Ichikawa T, Breakefield XO, Chiocca EA (2001) Improved helper virus-free packaging system for HSV amplicon vectors using an ICP27-deleted, oversized HSV-1 DNA in a bacterial artificial chromosome. Mol Ther 3:591–601
Zaupa C, Revol-Guyot V, Epstein AL (2003) Improved packaging system for generation of high levels non-cytotoxic HSV-1 amplicon vectors using Cre-loxP1 site-specific recombination to delete the packaging signals of defective helper genomes. Hum Gene Ther 14:1049–1063
Smith IL, Hardwicke MA, Sandri-Goldin RM (1992) Evidence that the herpes simplex virus immediate early protein ICP27 acts post-transcriptionally during infection to regulate gene expression. Virology 186:74–86
Krisky DM, Wolfe D, Goins WF, Marconi PC, Ramakrishnan R, Mata M, Rouse RJ, Fink DJ, Glorioso JC (1998) Deletion of multiple immediate-early genes from herpes simplex virus reduces cytotoxicity and permits long-term gene expression in neurons. Gene Ther 5:1593–1603
Kashima T, Vinters HV, Campagnoni AT (1995) Unexpected expression of intermediate filament protein genes in human oligodendroglioma cell lines. J Neuropathol Exp Neurol 54:23–31
Melendez ME, Aguirre AI, Baez MV, Bueno CA, Salvetti A, Jerusalinsky DA, Epstein AL (2013) Improvements in HSV-1-derived amplicon vectors for gene transfer, Chapter 1. In: Borrelli J, Giannini Y (eds) Advances in virus genomes research. Nova Science Publishers, Hauppauge, NY, pp 1–49
McGeoch DJ, Dalrymple MA, Davison AJ, Dolan A, Frame MC, McNab D, Perry LJ, Scott JE, Taylor P (1988) The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol 69:1531–15374
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Fraefel, C., Epstein, A.L. (2020). Preparation of Herpes Simplex Virus Type 1 (HSV-1)-Based Amplicon Vectors. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus . Methods in Molecular Biology, vol 2060. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9814-2_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9814-2_5
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9813-5
Online ISBN: 978-1-4939-9814-2
eBook Packages: Springer Protocols