1. Role of ERCC1 in removal of long non‐homologous tails during targeted homologous recombination.
- Author
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Adair, Gerald M., Rolig, Rhonda L., Moore‐Faver, Dana, Zabelshansky, Marina, Wilson, John H., and Nairn, Rodney S.
- Subjects
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HOMOLOGOUS recombination , *EXCISION repair , *DNA repair , *SACCHAROMYCES cerevisiae , *CELL lines , *CHO cell , *GENE targeting - Abstract
The XpF/Ercc1 structure‐specific endonuclease performs the 5′ incision in nucleotide excision repair and is the apparent mammalian counterpart of the Rad1/Rad10 endonuclease from Saccharomyces cerevisiae. In yeast, Rad1/Rad10 endonuclease also functions in mitotic recombination. To determine whether XpF/Ercc1 endonuclease has a similar role in mitotic recombination, we targeted the APRT locus in Chinese hamster ovary ERCC1+ and ERCC1− cell lines with insertion vectors having long or short terminal non‐homologies flanking each side of a double‐strand break. No substantial differences were evident in overall recombination frequencies, in contrast to results from targeting experiments in yeast. However, profound differences were observed in types of APRT+ recombinants recovered from ERCC1− cells using targeting vectors with long terminal non‐homologies—almost complete ablation of gap repair and single‐reciprocal exchange events, and generation of a new class of aberrant insertion/deletion recombinants absent in ERCC1+ cells. These results represent the first demonstration of a requirement for ERCC1 in targeted homologous recombination in mammalian cells, specifically in removal of long non‐homologous tails from invading homologous strands. [ABSTRACT FROM AUTHOR]
- Published
- 2000
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