Microbubbles for Nucleic Acid Delivery in Liver Using Mild Sonoporation

Methods Mol Biol. 2019:1943:377-387. doi: 10.1007/978-1-4939-9092-4_25.

Abstract

Ultrasound-mediated gene delivery is an interesting approach, which could help in increasing gene transfer in deep tissues. Moreover, it allows for performing experiments guided by the image to determine which elements are required. Microbubbles complexed with a eukaryotic expression cassette are excellent agents as they are responsive to ultrasounds and, upon oscillation, can destabilize membranes to enhance gene transfer. Here, we describe the preparation of positively charged microbubbles, plasmid free of antibiotic resistance marker, their combination and the conditions of ultrasound-mediated liver transfection post-systemic administration in mice. This association allowed us to obtain a superior liver gene expression at least over 8 months after a single injection.

Keywords: Cationic lipid; Gene expression; Lipoplex characterization; Microbubbles; Optical imaging; Ultrasound delivery; pFAR.

MeSH terms

  • Animals
  • Cell Membrane Permeability / radiation effects
  • Genetic Therapy / methods
  • HeLa Cells
  • Humans
  • Liver / cytology
  • Liver / metabolism
  • Liver / radiation effects
  • Mice
  • Mice, Inbred BALB C
  • Microbubbles*
  • Nucleic Acids / genetics
  • Transfection / methods*
  • Ultrasonic Waves*

Substances

  • Nucleic Acids