Enhanced cleavage of double-stranded DNA by artificial zinc-finger nuclease sandwiched between two zinc-finger proteins

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

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

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

Original languageEnglish
Pages (from-to)12257-12259
Number of pages3
JournalBiochemistry
Volume47
Issue number47
DOIs
Publication statusPublished - Nov 25 2008
Externally publishedYes

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Zinc Fingers
Zinc
DNA
Proteins
Micrococcal Nuclease
DNA Cleavage
Protein Binding
Fingers
Monomers
Binding Sites
Technology
Substrates
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Enhanced cleavage of double-stranded DNA by artificial zinc-finger nuclease sandwiched between two zinc-finger proteins. / Mineta, Yusuke; Okamoto, Tomoyuki; Takenaka, Kosuke; Doi, Norio; Aoyama, Yasuhiro; Sera, Takashi.

In: Biochemistry, Vol. 47, No. 47, 25.11.2008, p. 12257-12259.

Research output: Contribution to journalArticle

Mineta, Yusuke ; Okamoto, Tomoyuki ; Takenaka, Kosuke ; Doi, Norio ; Aoyama, Yasuhiro ; Sera, Takashi. / Enhanced cleavage of double-stranded DNA by artificial zinc-finger nuclease sandwiched between two zinc-finger proteins. In: Biochemistry. 2008 ; Vol. 47, No. 47. pp. 12257-12259.
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