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 language | English |
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Pages (from-to) | 12257-12259 |
Number of pages | 3 |
Journal | Biochemistry |
Volume | 47 |
Issue number | 47 |
DOIs | |
Publication status | Published - Nov 25 2008 |
Externally published | Yes |
ASJC Scopus subject areas
- Biochemistry