Protocol

Lipoplex and LPD Nanoparticles for In Vivo Gene Delivery

  1. Huang Leaf1,3
  1. 1 Division of Molecular Pharmaceutics, School of Pharmacy, University of North Carolina at Chapel Hill, North Carolina 27599, USA
  2. 2 Center for Pharmacogenetics, School of Pharmacy, University of Pittsburgh, Pennsylvania 15213, USA
  1. 3Corresponding author (leafh{at}pitt.edu)

INTRODUCTION

Lipoplex (cationic liposome-DNA complex) is formed via electrostatic interaction of anionic nucleic acids with cationic liposomes. A thin film of lipids is dried on the bottom of a glass tube and rehydrated in an aqueous solution. The resulting liposome suspension is passed through polycarbonate filters of desired pore size. This protocol also describes the preparation, physical properties, and biological activity of liposome-polycation-DNA (LPD) nanoparticles. The LPD nanoparticles contain a highly condensed DNA core surrounded by lipid bilayers with an average size of ~100 nm. The nanoparticle complex is injected into mice, and expression of the transfected DNA is monitored with an appropriate assay.

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