Abstract
Amplicons are defective, helper -dependent, herpes simplex virus type 1 (HSV-1 )-derived vectors. The main interest of these vectors as gene transfer tools stems from the fact that the amplicon vector genomes do not carry protein-encoding viral sequences . Consequently, they are completely safe for the host and non-toxic for the infected cells . Moreover, the complete absence of virus genes provides space to accommodate very large foreign DNA sequences, up to almost 150-kbp, the size of the virus genome . This large 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 . During 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 summarizes these two methodologies.
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Abbreviations
- A/H:
-
Amplicon/helper
- HC:
-
Helper -contaminated stocks
- HF:
-
Helper-free stocks
- PFU:
-
Plaque forming units
- TU:
-
Transducing units
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Fraefel, C., Marconi, P., Epstein, A.L. (2015). Herpes Simplex Virus Type 1 (HSV-1 )-Derived Amplicon Vectors for Gene Transfer and Gene Therapy . In: Lossi, L., Merighi, A. (eds) Neuronal Cell Death. Methods in Molecular Biology, vol 1254. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2152-2_21
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DOI: https://doi.org/10.1007/978-1-4939-2152-2_21
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