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- W2972029260 abstract "Skin’s exposure to UV radiation generates DNA damages, mended by Nucleotide Excision Repair (NER). Mutations in NER DNA damage recognition protein, Xeroderma Pigmentosum protein C (XPC), increases skin cancer incidence. The phenotype of XPC cells encompasses photosensitivity and accumulation of DNA lesions. Therefore, we aim to develop and execute screening methods to identify RNAi and chemical products reversing this phenotype. The screening procedure consists of subjecting XPC and normal fibroblasts to either libraries, UV irradiation then assessment of reversal. Hits are molecules inducing both increased viability post UV quantified by Presto Blue and repair of DNA lesions by ICC using Cell insight single cell analysis. Controls are non-treated cells either irradiated (negative controls) or UV protected (positive controls) via the modification of the plate covers blocking UV form desired spots. 24 hours post UV was chosen as readout time point after time varied UV dose-response curves and repair kinetics analysis. The applicability of chemical screen on large scale was confirmed by Z’ factor of 0.6 and 0.723 for the XPC and normal cells respectively. Moreover, chemical treatment will be performed pre and post UV to classify the effect as either preventative or therapeutic. For the siRNA screen, lowest toxicity and highest transfection efficiency were the criteria for choosing the transfection reagent, its ratio and transfection duration of 48 hours. We validated the screening methods using the controls which enabled the commencement of library screening currently being performed. After ruling out off-target effects and false positives, the hits will be tested on 3D skin models to mimic physiologic conditions as perspective." @default.
- W2972029260 created "2019-09-12" @default.
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- W2972029260 date "2019-09-01" @default.
- W2972029260 modified "2023-09-30" @default.
- W2972029260 title "535 RNA interference and Chemical-Based High Content Screening for the Normlaization of XPC Phenotype" @default.
- W2972029260 doi "https://doi.org/10.1016/j.jid.2019.07.450" @default.
- W2972029260 hasPublicationYear "2019" @default.
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