TY - JOUR
T1 - Multiple-turnover cleavage of double-stranded DNA by sandwiched zinc-finger nuclease.
AU - Mineta, Yusuke
AU - Okamoto, Tomoyuki
AU - Takenaka, Kosuke
AU - Doi, Norio
AU - Aoyama, Yasuhiro
AU - Sera, Takashi
N1 - Copyright:
This record is sourced from MEDLINE/PubMed, a database of the U.S. National Library of Medicine
PY - 2009
Y1 - 2009
N2 - 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.
AB - 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.
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U2 - 10.1093/nass/nrp140
DO - 10.1093/nass/nrp140
M3 - Article
C2 - 19749369
AN - SCOPUS:77952520353
SN - 1746-8272
SP - 279
EP - 280
JO - Nucleic acids symposium series (2004)
JF - Nucleic acids symposium series (2004)
IS - 53
ER -