Uptake and transfection efficiency of PEGylated cationic liposome-DNA complexes with and without RGD-tagging

Biomaterials. 2014 Jun;35(18):4996-5005. doi: 10.1016/j.biomaterials.2014.03.007. Epub 2014 Mar 21.


Steric stabilization of cationic liposome-DNA (CL-DNA) complexes is required for in vivo applications such as gene therapy. PEGylation (PEG: poly(ethylene glycol)) of CL-DNA complexes by addition of PEG2000-lipids yields sterically stabilized nanoparticles but strongly reduces their gene delivery efficacy. PEGylation-induced weakening of the electrostatic binding of CL-DNA nanoparticles to cells (leading to reduced uptake) has been considered as a possible cause, but experimental results have been ambiguous. Using quantitative live-cell imaging in vitro, we have investigated cell attachment and uptake of PEGylated CL-DNA nanoparticles with and without a custom synthesized RGD-peptide grafted to the distal ends of PEG2000-lipids. The RGD-tagged nanoparticles exhibit strongly increased cellular attachment as well as uptake compared to nanoparticles without grafted peptide. Transfection efficiency of RGD-tagged PEGylated CL-DNA NPs increases by about an order of magnitude between NPs with low and high membrane charge density (σM; the average charge per unit area of the membrane; controlled by the molar ratio of cationic to neutral lipid), even though imaging data show that uptake of RGD-tagged particles is only slightly enhanced by high σM. This suggests that endosomal escape and, as a result, transfection efficiency of RGD-tagged NPs is facilitated by high σM. We present a model describing the interactions between PEGylated CL-DNA nanoparticles and the anionic cell membrane which shows how the PEG grafting density and membrane charge density affect adhesion of nanoparticles to the cell surface.

Keywords: Gene therapy; Liposome; Live cell imaging; Nanoparticle; Polyethylene glycol; RGD peptide.

Publication types

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

MeSH terms

  • Animals
  • Cations / chemistry*
  • Cell Adhesion
  • Cell Line
  • DNA / chemistry*
  • Genetic Therapy
  • Lipids / chemistry
  • Liposomes / chemistry*
  • Mice
  • Microscopy, Electron
  • Nanoparticles / chemistry
  • Oligopeptides / chemistry*
  • Polyethylene Glycols / chemistry
  • Static Electricity
  • Transfection*


  • Cations
  • Lipids
  • Liposomes
  • Oligopeptides
  • Polyethylene Glycols
  • arginyl-glycyl-aspartic acid
  • DNA