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Test fonctionnel de mesure des activités enzymatiques de réparation de l'ADN par excision resynthèse sur support miniaturisé : mise au point et applications

Jean-François Millau 1
1 LAN - Laboratoire Lésions des Acides Nucléiques
SCIB - UMR E3 - Service de Chimie Inorganique et Biologique : DSM/INAC/SCIB
Abstract : DNA repair is a very important cellular mechanism involved in various genetic diseases and in carcinogenesis. DNA repair systems exhibit complementarities and interactions recently highlighted. However, this complexity is not taken into account by functional assays available nowadays.
Therefore we have designed a miniaturized in vitro assay allowing specific, functional and parallelized measurement of DNA repair activities. This test is an adaptation of an excision resynthesis assay to the microarray format.
We first setup and optimized the microarray fabrication, the data analysis and normalization, and the repair reaction. We then validated the assay and demonstrated that DNA repair activities measured were specific. Finally we demonstrated the potential of the assay in two different domains. We first phenotyped different cell types: the results showed the similarity of human fibroblasts repair profiles, and also the different repair abilities existing between fibroblast, keratinocyte, and peripheral blood mononuclear cells. Moreover we studied adaptation of cells to gamma irradiation. We demonstrated that DNA repair activities are involved in the radio-adaptation of cells .
Potential applications of this assay could be important in fundamental research as in applied one such as molecular screening, and in diagnosis.
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Submitted on : Friday, January 26, 2007 - 1:38:37 PM
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Jean-François Millau. Test fonctionnel de mesure des activités enzymatiques de réparation de l'ADN par excision resynthèse sur support miniaturisé : mise au point et applications. Biochimie [q-bio.BM]. Université Joseph-Fourier - Grenoble I, 2006. Français. ⟨tel-00126858⟩

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