SEQUENCE SELECTIVE DOUBLE-STRAND DNA CLEAVAGE BY PEPTIDE NUCLEIC-ACID (PNA) TARGETING USING NUCLEASE S1

被引:133
作者
DEMIDOV, V
FRANKKAMENETSKII, MD
EGHOLM, M
BUCHARDT, O
NIELSEN, PE
机构
[1] RUSSIAN ACAD SCI,INST MOLEC GENET,KURCHATOV SQ,MOSCOW 123182,RUSSIA
[2] HC ORSTED INST,DEPT ORGAN CHEM,DK-2100 COPENHAGEN 0,DENMARK
[3] PANUM INST,DEPT BIOCHEM B,DK-2200 COPENHAGEN N,DENMARK
关键词
D O I
10.1093/nar/21.9.2103
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A novel method for sequence specific double strand DNA cleavage using PNA (peptide nucleic acid) targeting is described. Nuclease Sl digestion of double stranded DNA gives rise to double strand cleavage at an occupied PNA strand displacement binding site, and under optimized conditions complete cleavage can be obtained. The efficiency of this cleavage is more than 1 0 fold enhanced when a tandem PNA site is targeted, and additionally enhanced if this site is in trans rather than in cis orientation. Thus in effect, the PNA targeting makes the single strand specific nuclease Sl behave like a pseudo restriction endonuclease.
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页码:2103 / 2107
页数:5
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