Enhanced Cleavage of Double-Stranded DNA by Artificial Zinc-Finger Nuclease Sandwiched between Two Zinc-Finger Proteins

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• 作者：Yusuke Mineta ; Tomoyuki Okamoto ; Kosuke Takenaka ; Norio Doi ; Yasuhiro Aoyama ; Takashi Sera
• 刊名：Biochemistry
• 出版年：2008
• 出版时间：November 25, 2008
• 年：2008
• 卷：47
• 期：47
• 页码：12257-12259
• 全文大小：143K
• 年卷期：v.47,no.47(November 25, 2008)
• ISSN：1520-4995

文摘

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.