Direct real-time molecular scale visualisation of the degradation of condensed DNA complexes exposed to DNase I

Nucleic Acids Res. 2003 Jul 15;31(14):4001-5. doi: 10.1093/nar/gkg462.

Abstract

The need to protect DNA from in vivo degradation is one of the basic tenets of therapeutic gene delivery and a standard test for any proposed delivery vector. The currently employed in vitro tests, however, presently provide no direct link between the molecular structure of the vector complexes and their success in this role, thus hindering the rational design of successful gene delivery agents. Here we apply atomic force microscopy (AFM) in liquid to visualise at the molecular scale and in real time, the effect of DNase I on generation 4 polyamidoamine dendrimers (G4) complexed with DNA. These complexes are revealed to be dynamic in nature showing a degree of mobility, in some cases revealing the addition and loss of dendrimers to individual complexes. The formation of the G4-DNA complexes is observed to provide a degree of protection to the DNA. This protection is related to the structural morphology of the formed complex, which is itself shown to be dependent on the dendrimer loading and the time allowed for complex formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry
  • DNA / metabolism*
  • DNA / ultrastructure
  • Deoxyribonuclease I / metabolism*
  • Macromolecular Substances
  • Microscopy, Atomic Force / methods
  • Polyamines / chemistry
  • Time Factors

Substances

  • Macromolecular Substances
  • Polyamines
  • DNA
  • Deoxyribonuclease I