Multiple-turnover cleavage of double-stranded DNA by sandwiched zinc-finger nuclease.

Yusuke Mineta, Tomoyuki Okamoto, Kosuke Takenaka, Norio Doi, Yasuhiro Aoyama, Takashi Sera

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)


To refine zinc-finger nuclease (ZFN) technology, we constructed a sandwiched ZFN, in which a DNA cleavage enzyme was sandwiched with two artificial zinc-finger proteins (AZPs). Because the sandwiched ZFN is designed to cleave the DNA between the two AZP-binding sites, the sandwiched ZFN is expected to bind preferentially to a DNA substrate rather than to cleavage products and thereby cleave it with multiple turnovers. To prove the concept, we sandwiched a staphylococcal nuclease (SNase), which cleaves DNA as a monomer, between two 3-finger AZPs. The AZP-sandwiched SNase cleaved large amounts of dsDNA site-specifically. Such multiple-turnover cleavage was not observed with control nucleases that possess a single AZP.

Original languageEnglish
Pages (from-to)279-280
Number of pages2
JournalNucleic acids symposium series (2004)
Issue number53
Publication statusPublished - 2009
Externally publishedYes

ASJC Scopus subject areas

  • Medicine(all)


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