A Robust Post-Insertion Method for the Preparation of Targeted Lipid Nanoparticles

Methods Mol Biol. 2025:2901:241-250. doi: 10.1007/978-1-0716-4394-5_18.

Abstract

Lipid nanoparticles (LNPs) have garnered significant attention as nonviral vectors for gene delivery due to their low immunogenicity and cytotoxicity. In addition, their surfaces can readily be functionalized with targeting moieties. Here we describe a simple and robust method for the preparation of targeted siRNA, mRNA, or DNA LNPs. After conjugating a targeting ligand to a PEG lipid, this binder-lipid conjugate is post-inserted into preformed LNPs. This process allows the LNPs to be easily decorated with one or various combinations of ligands.

Keywords: Binders; Gene delivery; Lipid nanoparticles (LNPs); Nanomedicine; Particle functionalization; Post-insertion; Specificity; Targeted delivery; Targeting ligand.

MeSH terms

  • DNA / chemistry
  • DNA / genetics
  • Gene Transfer Techniques
  • Humans
  • Lipids* / chemistry
  • Liposomes
  • Nanoparticles* / chemistry
  • Polyethylene Glycols / chemistry
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / genetics

Substances

  • Lipids
  • RNA, Small Interfering
  • Polyethylene Glycols
  • RNA, Messenger
  • Lipid Nanoparticles
  • DNA
  • Liposomes