Nucleic acid-mediated intracellular protein delivery by lipid-like nanoparticles

Biomaterials. 2014 Aug;35(24):6454-61. doi: 10.1016/j.biomaterials.2014.04.014. Epub 2014 May 13.

Abstract

Intracellular protein delivery has potential biotechnological and therapeutic application, but remains technically challenging. In contrast, a plethora of nucleic acid carriers have been developed, with lipid-based nanoparticles (LNPs) among the most clinically advanced reagents for oligonucleotide delivery. Here, we validate the hypothesis that oligonucleotides can serve as packaging materials to facilitate protein entrapment within and intracellular delivery by LNPs. Using two distinct model proteins, horseradish peroxidase and NeutrAvidin, we demonstrate that LNPs can yield efficient intracellular protein delivery in vitro when one or more oligonucleotides have been conjugated to the protein cargo. Moreover, in experiments with NeutrAvidin in vivo, we show that oligonucleotide conjugation significantly enhances LNP-mediated protein uptake within various spleen cell populations, suggesting that this approach may be particularly suitable for improved delivery of protein-based vaccines to antigen-presenting cells.

Keywords: Intracellular delivery; Lipid; Nanoparticles; Nucleic acid; Oligonucleotide; Protein delivery.

Publication types

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

MeSH terms

  • Animals
  • Avidin / metabolism*
  • Biophysical Phenomena
  • Carbocyanines / metabolism
  • DNA / metabolism
  • Drug Delivery Systems*
  • Electrophoresis, Polyacrylamide Gel
  • Female
  • HeLa Cells
  • Horseradish Peroxidase / metabolism*
  • Humans
  • Immunohistochemistry
  • Intracellular Space / metabolism*
  • Lipids / chemistry*
  • Mice, Inbred C57BL
  • Nanoparticles / chemistry*
  • Nucleic Acids / metabolism*
  • Oligonucleotides / metabolism
  • Spectroscopy, Near-Infrared
  • Spleen / metabolism

Substances

  • Carbocyanines
  • Lipids
  • Nucleic Acids
  • Oligonucleotides
  • cyanine dye 5
  • neutravidin
  • Avidin
  • DNA
  • Horseradish Peroxidase