Abstract
Genetic alterations affecting nucleotide excision repair, the most versatile DNA-repair mechanism responsible for removal of bulky DNA adducts including ultraviolet (UV) light-induced DNA lesions, may result in the rare, recessively inherited autosomal syndromes xeroderma pigmentosum (XP), Cockayne syndrome (CS), or trichothiodystrophy (TTD). Classical approaches such as somatic cell fusions or microinjection assays have formalized the genetic complexity of these related but clinically distinct syndromes, and contributed to the determination of seven, five, and three complementation groups for XP, CS, and TTD, respectively. XP patients are highly susceptible to photoinduced cutaneous cancers of epidermal origin. To better study the responses to UV irradiation of XP keratinocytes, and to objectively determine the extent to which cutaneous gene therapy may be realized, we set up experimental procedures adapted to ex vivo genetic complementation of keratinocytes from XP patients. We provide here detailed rationales and procedures for these approaches.
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Acknowledgments
MF and VB are equal contributors. This work was supported by grants from the Association pour la Recherche contre le Cancer (ARC, contract 9500 to T. M.; ARC, contract 5765 to A. S.), the Association Française contre les Myopathies (AFM, to T. M.), the Fondation de l’Avenir and funds from Centre National de la Recherche Scientifique (CNRS). MF is supported by Lóréal. We gratefully acknowledge Dr. F. Bernard for expert advice concerning skin reconstruction in vitro.
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Fréchet, M., Bergoglio, V., Chevallier-Lagente, O., Sarasin, A., Magnaldo, T. (2006). Complementation Assays Adapted for DNA Repair-Deficient Keratinocytes. In: Henderson, D.S. (eds) DNA Repair Protocols. Methods in Molecular Biology™, vol 314. Humana Press. https://doi.org/10.1385/1-59259-973-7:009
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DOI: https://doi.org/10.1385/1-59259-973-7:009
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