Complementation assays adapted for DNA repair-deficient keratinocytes

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 314; p. 9
• Main Authors: Fréchet, Mathilde, Bergoglio, Valérie, Chevallier-Lagente, Odile, Sarasin, Alain, Magnaldo, Thierry
• Format: Journal Article
• Language: English
• Published: United States 2006
• Subjects: DNA - radiation effects; DNA Repair - genetics; DNA Replication; Genetic Complementation Test - methods; Genetic Complementation Test - standards; Genetic Therapy; Genotype; Humans; Keratinocytes - pathology; Keratinocytes - radiation effects; Phenotype; Retroviridae - genetics; Skin Diseases - genetics; Skin Diseases - pathology; Skin Diseases - therapy; Transduction, Genetic; Ultraviolet Rays; Xeroderma Pigmentosum - genetics; Xeroderma Pigmentosum - pathology; Xeroderma Pigmentosum - therapy
• ISSN: 1064-3745
• DOI: 10.1385/1-59259-973-7:009

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.