Abstract
We report the design and optimisation of novel oligonucleotide substrates for a sensitive fluorescence assay for high-throughput screening and functional studies of the DNA repair enzyme, XPF-ERCC1, with a view to accelerating inhibitor and drug discovery.
Highlights
We report the design and optimisation of novel oligonucleotide substrates for a sensitive fluorescence assay for high-throughput screening and functional studies of the DNA repair enzyme, XPFERCC1, with a view to accelerating inhibitor and drug discovery
XPF-ERCC1 is involved in a number of DNA repair pathways including nucleotide excision repair (NER), homologous recombination (HR) and interstrand crosslinks (ICLs) repair, playing a critical role in the repair-response to a number of DNA damaging agents used in cancer therapy.[18]
Full-length XPF-ERCC1 complex was purified from insect cells using a baculovirus expression system [Fig. S1A (ESI†)]
Summary
We report the design and optimisation of novel oligonucleotide substrates for a sensitive fluorescence assay for high-throughput screening and functional studies of the DNA repair enzyme, XPFERCC1, with a view to accelerating inhibitor and drug discovery. McHugh, Tom Brown et al Optimised oligonucleotide substrates to assay XPF-ERCC1 nuclease activity for the discovery of DNA repair inhibitors
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