PMID: 8960128Dec 2, 1996Paper

Long-term complementation of DNA repair deficient human primary fibroblasts by retroviral transduction of the XPD gene

Mutation Research
X QuillietM Mezzina

Abstract

Due to their limited life time in culture and their relative resistance to DNA transfection, primary fibroblasts derived from UV-hypersensitive patients could not be used for cloning DNA repair gene and studying stable complementation with wild-type DNA repair genes. Primary cells were only used for complementation analysis after transient expression through cell fusion. DNA microinjection and transfection. We report the retroviral-mediated highly efficient transfer and stable expression of XPD/ERCC2 gene in fibroblast strains from eight different patients using the LXPDSN retroviral vector. Cells derived from skin biopsies of xeroderma pigmentosum and trichothiodystrophy patients were incubated with vector-containing suspension and selected with the neomycin-analog G418. LXPDSN vector specifically complemented cells belonging to the XP-D group. Long-term reversion of repair-deficient phenotype, monitored by UV survival and UDS analysis, has been achieved in these diploid fibroblasts. We demonstrate this methodology is a powerful tool to study phenotypic reversion of nucleotide excision repair-deficient cells such as cellular DNA repair properties and we suggest that it may be used to study other cellular parameters (cell cycle...Continue Reading

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Citations

Apr 21, 1999·International Journal of Cancer. Journal International Du Cancer·G GigliaA Sarasin
Nov 13, 2002·Mutation Research·Thierry Magnaldo, Alain Sarasin
Nov 10, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Emilie WarrickThierry Magnaldo
Jul 5, 2001·Proceedings of the National Academy of Sciences of the United States of America·F BernerdT Magnaldo
Jun 24, 1997·Proceedings of the National Academy of Sciences of the United States of America·C AhrensJ Krutmann
Aug 5, 1997·Proceedings of the National Academy of Sciences of the United States of America·E M TaylorA R Lehmann
May 20, 2000·Biotechnology & Genetic Engineering Reviews·S BevanI A Mckay
Jul 19, 2003·Human Gene Therapy·Catherine Arnaudeau-BégardThierry Magnaldo
Oct 12, 2013·International Journal of Molecular Sciences·Sophie RouanetThierry Magnaldo
Jan 15, 2005·Cancer Gene Therapy·Melissa Gava ArmeliniCarlos Frederico Martins Menck

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