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Human artificial chromosome (HAC) vectors are an important gene transfer system for expression and complementation studies. We describe a significant advance in HAC technology using infectious herpes simplex virus type 1 (HSV-1) amplicon vectors for delivery. This highly efficient method has allowed gene-expressing HACs to be established in glioma-, kidney- and lung-derived cells. We also developed an HSV-1 hypoxanthine phosphoribosyltransferase (HPRT) HAC vector, which generated functional HPRT-expressing HACs that complemented the genetic deficiency in human cells. The transduction efficiency of the HSV-1 HAC amplicons is several orders of magnitude higher than lipofection-mediated delivery. Studies on HAC stability between cell types showed important differences that have implications for HAC development and gene expression in human cells. This is the first report of establishing gene-expressing HACs in human cells by using an efficient, high-capacity viral vector and by identifying factors that are involved in cell-type-specific HAC instability. The work is a significant advance for HAC technology and the development of HAC gene expression systems in human cells.

Original publication

DOI

10.1038/sj.embor.7400768

Type

Journal article

Journal

EMBO Rep

Publication Date

09/2006

Volume

7

Pages

911 - 918

Keywords

Autoantigens, Cell Line, Centromere Protein A, Chromosomal Instability, Chromosomal Proteins, Non-Histone, Chromosomes, Artificial, Human, Clone Cells, Gene Transfer Techniques, Genetic Vectors, Green Fluorescent Proteins, Herpesvirus 1, Human, Humans, Mitosis, Transfection