Faculty Articles
Analysis of actively transcribed DNA repair using a transfection-based system
Publication Title
Methods in molecular biology
Publisher
Humana Press
ISSN
1064-3745
Publication Date
1-1-2014
Keywords
Animals, Bacteriophages, Cell Line, DNA Repair, Genes, Reporter, Humans, Luciferases, Firefly, Plasmids, Transfection, Ultraviolet Rays
Abstract
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 as an exemplar UV irradiation as a source of damage to a luciferase reporter plasmid. Through measurement of the activity of a successfully transcribed and translated 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. Since 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, a subset of the overall nucleotide excision repair pathway that specifically targets transcribed gene sequences.
DOI
10.1007/978-1-62703-739-6_37
Volume
1105
First Page
533
Last Page
550
Disciplines
Medicine and Health Sciences | Pharmacy and Pharmaceutical Sciences
NSUWorks Citation
Latimer, Jean Johanna, "Analysis of actively transcribed DNA repair using a transfection-based system" (2014). Faculty Articles. 22.
https://nsuworks.nova.edu/hpd_corx_facarticles/22
