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• W2034383318 endingPage "179" @default.
• W2034383318 startingPage "170" @default.
• W2034383318 abstract "Targeted gene repair, a form of oligonucleotide-directed mutagenesis, employs end-modified single-stranded DNA oligonucleotides to mediate single-base changes in chromosomal DNA. In this work, we use a specific 72-mer to direct the repair of a mutated eGFP gene stably integrated in the genome of DLD-1 cells. Corrected cells express eGFP that can be identified and quantitated by FACS. The repair of this mutant gene is dependent on the presence of a specifically designed oligonucleotide and the frequency with which the mutation is reversed is affected by the induction of DNA damage. We used hydroxyurea, VP16 (etoposide), and thymidine to modulate the rate of DNA replication through the stalling of the replication forks or the introduction of lesions. Addition of hydroxyurea or VP16 before the electroporation of the oligonucleotide, results in an accumulation of double-strand breaks (DSB) whose repair is facilitated by either nonhomologous end joining (NHEJ) or homologous recombination (HR). The addition of thymidine results in DNA damage within replication forks, damage that is repaired through the process of homologous recombination. Our data suggest that gene repair activity is elevated when DNA damage induces or activates the homologous recombination pathway." @default.
• W2034383318 created "2016-06-24" @default.
• W2034383318 creator A5024782604 @default.
• W2034383318 creator A5051654097 @default.
• W2034383318 creator A5081138565 @default.
• W2034383318 date "2004-10-01" @default.
• W2034383318 modified "2023-09-26" @default.
• W2034383318 title "Enhanced oligonucleotide-directed gene targeting in mammalian cells following treatment with DNA damaging agents" @default.
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• W2034383318 doi "https://doi.org/10.1016/j.yexcr.2004.06.021" @default.
• W2034383318 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/15383324" @default.
• W2034383318 hasPublicationYear "2004" @default.
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