Intracellular delivery of nanoparticles and DNAs by IR9 cell-penetrating peptides

PLoS One. 2013 May 28;8(5):e64205. doi: 10.1371/journal.pone.0064205. Print 2013.

Abstract

Cell-penetrating peptides (CPPs) comprised of basic amino residues are able to cross cytoplasmic membranes and are able to deliver biologically active molecules inside cells. However, CPP/cargo entrapment in endosome limits biomedical utility as cargoes are destroyed in the acidic environment. In this study, we demonstrate protein transduction of a novel CPP comprised of an INF7 fusion peptide and nona-arginine (designated IR9). IR9 noncovalently interacts with quantum dots (QDs) and DNAs to form stable IR9/QD and IR9/DNA complexes which are capable of entering human A549 cells. Zeta-potentials were a better predictor of transduction efficiency than gel shift analysis, emphasizing the importance of electrostatic interactions of CPP/cargo complexes with plasma membranes. Mechanistic studies revealed that IR9, IR9/QD and IR9/DNA complexes may enter cells by endocytosis. Further, IR9, IR9/QD and IR9/DNA complexes were not cytotoxic at concentrations below 30, 5 and 20.1 µM, respectively. Without labor intensive production of fusion proteins from prokaryotes, these results indicate that IR9 could be a safe carrier of genes and drugs in biomedical applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Death / drug effects
  • Cell-Penetrating Peptides / pharmacology*
  • DNA / metabolism*
  • Drug Delivery Systems*
  • Endocytosis / drug effects
  • Flow Cytometry
  • Humans
  • Intracellular Space / drug effects
  • Intracellular Space / metabolism*
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Nanoparticles / chemistry*
  • Plasmids / metabolism
  • Quantum Dots
  • Static Electricity

Substances

  • Cell-Penetrating Peptides
  • DNA

Grants and funding

The financial support of this study was provided by the Postdoctoral Fellowship NSC 101-2811-B-259-001 (to Liu BR) and the Grant Number NSC 101-2320-B-259-002-MY3 from the National Science Council of Taiwan (to Lee HJ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.