Title

Analysis of DNA repair using transfection-based host cell reactivation

ISBN or ISSN

1064-3745

Publication Title

Methods in molecular biology

Volume

291

Publication Date / Copyright Date

1-1-2005

First Page

321

Last Page

335

Publisher

Humana Press

DOI Number

10.1385/1-59259-840-4:321

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 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.

Disciplines

Medicine and Health Sciences | Pharmacy and Pharmaceutical Sciences

Keywords

Animals, Base Pair Mismatch, Cells, Cultured, DNA, DNA Damage, DNA Repair, Genes, Reporter, Humans, Luciferases, Plasmids, Pyrimidine Dimers, Transfection

This document is currently not available here.

Peer Reviewed

Find in your library

Share

 
COinS