Sequence-specific targeting of duplex DNA by peptide nucleic acids via triplex strand invasion

Methods. 2001 Feb;23(2):108-22. doi: 10.1006/meth.2000.1112.

Abstract

Because of a set of exceptional chemical, physical, and biological properties, polyamide or peptide nucleic acids (PNAs) hold a distinctive position among various synthetic ligands designed for DNA-targeting purposes. Cationic pyrimidine PNAs (cpyPNAs) represent a special group of PNAs, which effectively form strand invasion triplexes with double-stranded DNA (dsDNA) also known as P-loops. Extraordinary stability of the invasion triplexes and high sequence specificity of their formation combined with local opening of the DNA double helix within the P-loops make these complexes very attractive for sequence-specific manipulation with dsDNA. Important for applications is the fact that the discrimination between correct and mismatched binding sites in dsDNA by cpyPNAs is a nonequilibrium, kinetically controlled process. Therefore, a careful choice of experimental conditions that are optimal for the kinetic discrimination of correct versus mismatched cpyPNA binding is crucial for sequence-specific recognition of dsDNA by cpyPNAs. The experimental and theoretical data presented make it possible to select those solution parameters and cpyPNA constructions that are most favorable for sequence specificity without compromising the affinity of dsDNA targeting.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Pair Mismatch
  • Binding Sites
  • DNA / chemistry*
  • DNA / metabolism
  • Escherichia coli / genetics
  • Genetic Techniques*
  • Humans
  • Hydrogen-Ion Concentration
  • Kinetics
  • Models, Statistical
  • Peptide Nucleic Acids / chemistry
  • Peptide Nucleic Acids / metabolism
  • Plasmids / metabolism
  • Pyrimidines / chemistry
  • Saccharomyces cerevisiae / genetics
  • Time Factors

Substances

  • Peptide Nucleic Acids
  • Pyrimidines
  • triplex DNA
  • DNA