Cationic Dendrimers for siRNA Delivery: An Overview of Methods for In Vitro/In Vivo Characterization

Methods Mol Biol. 2021:2282:209-244. doi: 10.1007/978-1-0716-1298-9_14.

Abstract

This chapter reviews the different techniques for analyzing the chemical-physical properties, transfection efficiency, cytotoxicity, and stability of covalent cationic dendrimers (CCDs) and self-assembled cationic dendrons (ACDs) for siRNA delivery in the presence and absence of their nucleic cargos. On the basis of the reported examples, a standard essential set of techniques is described for each step of a siRNA/nanovector (NV) complex characterization process: (1) analysis of the basic chemical-physical properties of the NV per se; (2) characterization of the morphology, size, strength, and stability of the siRNA/NV ensemble; (3) characterization and quantification of the cellular uptake and release of the siRNA fragment; (4) in vitro and (5) in vivo experiments for the evaluation of the corresponding gene silencing activity; and (6) assessment of the intrinsic toxicity of the NV and the siRNA/NV complex.

Keywords: Cationic dendrimer; Experimental characterization; PAMAM dendrimers; Self-assembling nanocarriers; siRNA delivery.

Publication types

  • Review

MeSH terms

  • Animals
  • Cations
  • Cell Line
  • Dendrimers / chemistry*
  • Dendrimers / metabolism
  • Dendrimers / toxicity
  • Humans
  • RNA Interference*
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / genetics*
  • RNA, Small Interfering / metabolism
  • RNAi Therapeutics*
  • Transfection*

Substances

  • Cations
  • Dendrimers
  • PAMAM Starburst
  • RNA, Small Interfering