Correction of Frameshift Mutations with Tailed Duplex DNAs

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  • Morita Yukiko
    Faculty of Pharmaceutical Sciences, Hokkaido University
  • Tsuchiya Hiroyuki
    Faculty of Pharmaceutical Sciences, Hokkaido University
  • Harashima Hideyoshi
    Faculty of Pharmaceutical Sciences, Hokkaido University Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST)
  • Kamiya Hiroyuki
    Faculty of Pharmaceutical Sciences, Hokkaido University Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST) Graduate School of Science and Engineering, Ehime University

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Tailed duplex (TD) DNAs, prepared by annealing an oligonucleotide to a several-hundred-base single-stranded (ss) DNA fragment, correct a base-substitution mutation with high efficiency. In the present study, the abilities of TD fragments to correct single-base insertion and deletion mutations were examined, using hygromycin-resistance and enhanced green fluorescent protein fusion (Hyg-EGFP) genes inactivated by +G and −C frameshift mutations. The 5′-TD and 3′-TD DNA fragments were co-transfected with plasmid DNA containing the inactivated Hyg-EGFP gene into CHO-K1 cells, and the gene correction efficiencies were determined by introducing the plasmid DNA recovered from the transfected cells into Escherichia coli cells. In contrast to their efficiencies for the substitution mutation, the gene correction abilities of the TD fragments were relatively low. The correction efficiencies by the TD fragments were apparently higher than that by a ss DNA fragment, one of the DNA fragments employed for gene correction. These results suggest that the TD fragments have the potential to correct frameshift mutations, although further improvement is required.

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