Episomal transgene expression in pluripotent stem cells.

Journal article

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.

Authors: Lufino, M; Popplestone, A; Cowley, S; Edser, P; James, W

• Journal: Methods Mol Biol
• Volume: 767
• Pages: 369-387
• Publication date: 2011-01-01
• DOI: 10.1007/978-1-61779-201-4_27
• EISSN: 1940-6029
• ISSN: 1064-3745
• Language: English
• Keywords: Cell Line; Mice; Centrifugation, Density Gradient; Herpesvirus 1, Human; Dopaminergic Neurons; Cell Differentiation; Filtration; Humans; Pluripotent Stem Cells; Titrimetry; Clone Cells; Gene Expression; Transgenes; Virus Assembly; Embryonic Stem Cells; Genetic Vectors; Gene Transfer Techniques; Virion; Plasmids; Animals; DNA