Document Detail

Analysis of DNA double-strand break repair by nonhomologous end joining in cell-free extracts from mammalian cells.
MedLine Citation:
PMID:  15502235     Owner:  NLM     Status:  MEDLINE    
Double-strand breaks (DSBs) in genomic DNA are induced by ionizing radiation or radiomimetic drugs, but they also occur spontaneously during the cell cycle at quite significant frequencies. In vertebrate cells, nonhomologous DNA end joining (NHEJ) is considered the major pathway of DSB repair. NHEJ is able to rejoin two broken DNA termini directly end-to-end irrespective of sequence and structure. Genetic studies in various radiosensitive and DSB repair-deficient hamster cell lines have yielded insights into the factors involved in NHEJ. Studies in cell-free systems derived from Xenopus eggs and mammalian cells have allowed the dissection of the underlying mechanisms. In the present chapter, we describe a protocol for the preparation of whole cell extracts from mammalian cells and a plasmid-based in vitro assay that permits the easy analysis of the efficiency and fidelity of DSB repair via NHEJ in different cell types.
Petra Pfeiffer; Elke Feldmann; Andrea Odersky; Steffi Kuhfittig-Kulle; Wolfgang Goedecke
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  291     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2005  
Date Detail:
Created Date:  2004-10-25     Completed Date:  2005-02-22     Revised Date:  2006-05-01    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  351-71     Citation Subset:  IM    
Institute of Genetics, University of Essen, Germany.
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MeSH Terms
Base Sequence
CHO Cells
Cell Extracts / chemistry
Cell-Free System*
DNA / analysis,  chemistry
DNA Damage
DNA Repair*
Molecular Sequence Data
Plasmids / genetics
Recombination, Genetic*
Reg. No./Substance:
0/Cell Extracts; 9007-49-2/DNA

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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