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Sequence selective double strand DNA cleavage by peptide nucleic acid (PNA) targeting using nuclease S1
V Demidov1, M D Frank-Kamenetskii, M Egholm
1Institute of Molecular Genetics, Russian Academy of Sciences, Moscow.
Abstract:
A novel method for sequence specific double strand DNA cleavage using PNA (peptide nucleic acid) targeting is described. Nuclease S1 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 10 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 S1 behave like a pseudo restriction endonuclease.