Unscheduled DNA synthesis: the clinical and functional assay for global genomic DNA nucleotide excision repair.

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 2014 ;1105:511-32

Department of Pharmaceutical Sciences, Nova Southeastern University, 3301 College Avenue, Fort Lauderdale-Davie, FL, 33314-7796, USA,

The unscheduled DNA synthesis (UDS) assay measures the ability of a cell to perform global genomic nucleotide excision repair (NER). This chapter provides instructions for the application of this technique by creating 6-4 photoproducts and pyrimidine dimers using UV-C irradiation. This procedure is designed specifically for quantification of the 6-4 photoproducts. Repair is quantified by the amount of radioactive thymidine incorporated during repair synthesis after this insult, and radioactivity is evaluated by grain counting after autoradiography. The results are used to clinically diagnose human DNA repair deficiency disorders and provide a basis for investigation of repair deficiency in human tissues or tumors. No other functional assay is available that directly measures the capacity to perform NER on the entire genome without the use of specific antibodies. Since live cells are required for this assay, explant culture techniques must be previously established. 

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-62703-739-6_36DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4751080PMC
November 2014
158 Reads
7 Citations
1.300 Impact Factor

Publication Analysis

Top Keywords

6-4 photoproducts
8
repair deficiency
8
functional assay
8
global genomic
8
nucleotide excision
8
unscheduled dna
8
excision repair
8
dna synthesis
8
repair
7
hcr discussed
4
discussed chapter
4
radioactivity evaluated
4
counting autoradiography
4
diagnose human
4
human dna
4
dna repair
4
clinically diagnose
4
autoradiography clinically
4
grain counting
4
insult radioactivity
4

References

(Supplied by CrossRef)

LH Thompson et al.
1998

RD Wood et al.
Science 2001

E Reed et al.
Cancer Treat Rev 1998

M Kaneko et al.
Cancer Res 1980

BS Andersson et al.
Cancer Chemother Pharmacol 1996

MP Gamesik et al.
Curr Pharm Des 1999

LHF Mullenders et al.
J Photochem Photobiol B 2001

D Covertey et al.
Nature 1991

JC Huang et al.
Proc Natl Acad Sci USA 1992

KK Shivji et al.
Cell 1992

L Grossman et al.
J Biol Chem 1993

Similar Publications