核酸内切酶
DNA
高通量筛选
分子生物学
劈理(地质)
化学
DNA修复
生物
生物化学
断裂(地质)
古生物学
作者
Claire McWhirter,Michael Tonge,Helen Plant,Ian M. Hardern,J. Willem M. Nissink,Stephen T. Durant
标识
DOI:10.1177/1087057113476551
摘要
Flap endonuclease-1 (FEN1) is a highly conserved metallonuclease and is the main human flap endonuclease involved in the recognition and cleavage of single-stranded 5' overhangs from DNA flap structures. The involvement of FEN1 in multiple DNA metabolism pathways and the identification of FEN1 overexpression in a variety of cancers has led to interest in FEN1 as an oncology target. In this article, we describe the development of a 1536-well high-throughput screening assay based on the change in fluorescence polarization of a FEN1 DNA substrate labeled with Atto495 dye. The assay was subsequently used to screen 850 000 compounds from the AstraZeneca compound collection, with a Z' factor of 0.66 ± 0.06. Hits were followed up by IC50 determination in both a concentration-response assay and a technology artifact assay.
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