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 |
|---|---|
| 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 |
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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 journal › Article
}
TY - JOUR
T1 - Enhanced cleavage of double-stranded DNA by artificial zinc-finger nuclease sandwiched between two zinc-finger proteins
AU - Mineta, Yusuke
AU - Okamoto, Tomoyuki
AU - Takenaka, Kosuke
AU - Doi, Norio
AU - Aoyama, Yasuhiro
AU - Sera, Takashi
PY - 2008/11/25
Y1 - 2008/11/25
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=56749128115&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=56749128115&partnerID=8YFLogxK
U2 - 10.1021/bi801800k
DO - 10.1021/bi801800k
M3 - Article
C2 - 18980382
AN - SCOPUS:56749128115
VL - 47
SP - 12257
EP - 12259
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 47
ER -