Two-photon fluorescence cross-correlation spectroscopy as a potential tool for high-throughput screening of DNA repair activity | Zendy

Maddalena Collini | Zendy

Open Access

Two-photon fluorescence cross-correlation spectroscopy as a potential tool for high-throughput screening of DNA repair activity

Author(s) -

Maddalena Collini

Publication year - 2005

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gni166

Subject(s) - dna glycosylase , biology , uracil dna glycosylase , dna , dna repair , fluorescence cross correlation spectroscopy , fluorescence spectroscopy , dna damage , mutyh , biochemistry , microbiology and biotechnology , fluorescence , physics , quantum mechanics

Several lines of evidence indicate that differences in DNA repair capacity are an important source of variability in cancer risk. However, traditional assays for measurement of DNA repair activity in human samples are laborious and time-consuming. DNA glycosylases are the first step in base excision repair of a variety of modified DNA bases. Here, we describe the development of a new sensitive DNA glycosylase assay based on fluorescence cross-correlation spectroscopy (FCCS) with two-photon excitation. FCCS was applied to the measurement of uracil DNA glycosylase activity of human cell extracts and validated by comparison with standard gel electrophoresis assay. Our results indicate that FCCS can be adapted to efficient assays for DNA glycosylase activity in protein extracts from human cells. This method has a potential for the development of automated screening of large number of samples.

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