Analysis of DNA repair using transfection-based host cell reactivation.

Authors:
Dr. Jean J Latimer, Ph.D.
Dr. Jean J Latimer, Ph.D.
Nova Southeastern University AutoNation Cancer Institute
Director, AutoNation Institute for Breast Cancer
Breast cancer
Fort Lauderdale, Florida | United States

Methods Mol Biol 2005 ;291:321-35

Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, PA, USA.

Host cell reactivation (HCR) is a transfection-based assay in which intact cells repair damage localized to exogenous DNA. This chapter provides instructions for the application of this technique using UV irradiation as a source of damage to a luciferase reporter plasmid. Through measurement of the activity of a reporter enzyme, the amount of damaged plasmid that a cell can "reactivate" or repair and express can be quantitated. Different DNA repair pathways can be analyzed by this technique by damaging the reporter plasmid in different ways. Because it involves repair of a transcriptionally active gene, when applied to UV damage the HCR assay measures the capacity of the host cells to perform transcription-coupled repair (TCR), a subset of the overall nucleotide excision repair pathway that specifically targets transcribed gene sequences.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4860737PMC
http://dx.doi.org/10.1385/1-59259-840-4:321DOI Listing
February 2005
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27 Citations
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