Faculty Articles
Analysis of actively transcribed DNA repair using a transfection-based system
ISBN or ISSN
1064-3745
Publication Title
Methods in molecular biology
Volume
1105
Publication Date / Copyright Date
1-1-2014
First Page
533
Last Page
550
Publisher
Humana Press
DOI Number
10.1007/978-1-62703-739-6_37
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.
Disciplines
Medicine and Health Sciences | Pharmacy and Pharmaceutical Sciences
Keywords
Animals, Bacteriophages, Cell Line, DNA Repair, Genes, Reporter, Humans, Luciferases, Firefly, Plasmids, Transfection, Ultraviolet Rays
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
