Effects of target sequence and sense versus anti-sense strands on gene correction with single-stranded DNA fragments

Hiroyuki Kamiya, Masayuki Uchiyama, Yoshimichi Nakatsu, Teruhisa Tsuzuki, Hideyoshi Harashima

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Abstract

The correction of an inactivated hygromycin resistance and enhanced green fluorescent protein (Hyg-EGFP) fusion gene by a several hundred-base single-stranded (ss) DNA fragment has been reported. In this study, the effectiveness of this type of gene correction was examined for various positions in the rpsL gene. Sense and anti-sense ssDNA fragments were prepared, and the gene correction efficiencies were determined by co-introduction of the target plasmid containing the gene with the ssDNA fragments. The gene correction efficiency varied (0.8-9.3%), depending on target positions and sense/anti-sense strands. Sense ssDNA fragments corrected the target gene with equal or higher efficiencies as compared to their anti-sense counterparts. The target positions corrected with high efficiency by the sense fragments also tended to be corrected efficiently by the anti-sense fragments. These results suggest that the sense ssDNA fragments are useful for the correction of mutated genes. The variation in the correction efficiency may depend on the sequence of the target position in double-stranded DNA.

Original languageEnglish
Pages (from-to)431-436
Number of pages6
JournalJournal of biochemistry
Volume144
Issue number4
DOIs
Publication statusPublished - Oct 1 2008

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All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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