Sequence selective double strand DNA cleavage by peptide nucleic acid (PNA) targeting using nuclease S1

V Demidov, M D Frank-Kamenetskii, O Buchardt, Peter E. Nielsen

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.

Original languageEnglish
JournalNucleic Acids Research
Volume21
Issue number9
Pages (from-to)2103-7
Number of pages5
ISSN0305-1048
Publication statusPublished - 11 May 1993

Keywords

  • Base Sequence
  • DNA/metabolism
  • Genetic Techniques
  • Kinetics
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides/chemical synthesis
  • Single-Strand Specific DNA and RNA Endonucleases/metabolism
  • Substrate Specificity

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