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Herpes simplex type 1 (HSV-1) amplicon vectors possess a number of features that make them excellent vectors for the delivery of transgenes into stem cells. HSV-1 amplicon vectors are capable of efficiently transducing both dividing and nondividing cells and since the virus is quite large, 152 kb, it is of sufficient size to allow for incorporation of entire genomic DNA loci with native promoters. HSV-1 amplicon vectors can also be used to incorporate and deliver to cells a variety of sequences that allow extrachromosomal retention. These elements offer advantages over integrating vectors as they avoid transgene silencing and insertional mutagenesis. The construction of amplicon vectors carrying extrachromosomal retention elements, their packaging into HSV-1 viral particles, and the use of HSV-1 amplicons for stem cell transduction will be described.

Original publication

DOI

10.1007/978-1-61779-201-4_27

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2011

Volume

767

Pages

369 - 387

Keywords

Animals, Cell Differentiation, Cell Line, Centrifugation, Density Gradient, Clone Cells, DNA, Dopaminergic Neurons, Embryonic Stem Cells, Filtration, Gene Expression, Gene Transfer Techniques, Genetic Vectors, Herpesvirus 1, Human, Humans, Mice, Plasmids, Pluripotent Stem Cells, Titrimetry, Transgenes, Virion, Virus Assembly