Chapter 4
Assessing Nucleic Acid: Cationic Nanoparticle Interaction
for Gene Delivery
Moganavelli Singh
Abstract
The assessment of the efficient binding between a nucleic acid and its associated nanoparticle is crucial for
gene delivery. Emerging from the extensive search for versatile gene carriers, are complexes formed between
nucleic acids and nonviral nanocarriers that promise to be viable alternatives to the predominantly viralbased gene delivery vehicles. However, much is still to be known about the exact structure and physicochemical properties of such nanocomplexes. This chapter will concentrate on cationic lipid, polymer, and
functionalized metal nanoparticles and their interaction with nucleic acids by direct conjugation or electrostatic interaction. Methods commonly employed to evaluate the nature and extent of nucleic acid interactions with cationic nanocarriers, such a nucleic acid binding, nuclease protection, and dye displacement
assays will be described. In addition, the ultrastructural morphology, size, and zeta potential of these
nanocomplexes, which are crucial for their cellular uptake and intracellular trafficking, will be assessed
using electron microscopy, fluorescent detection, and nanoparticle tracking analysis (NTA). These assays
have the ability to visualize and quantify the interaction and can also be used to complement each other, in
addition to providing confirmation of the formation of the relevant nanocomplexes.
Key words Nucleic acids, Nanoparticles, Nanocarriers, Interaction, Complexes, Gene delivery
1 Introduction
The exhaustive search for safe and versatile gene delivery systems
has produced an array of different systems to date, ranging from the
popular viral systems to lipids, polymers, and inorganic nanoparticles such as gold, selenium, silver, iron oxide, and platinum, to
name a few. Important to the use of these carrier systems are
complexes formed between the nucleic acid and the nanocarrier.
The exact structure and properties of such nanocomplexes is crucial
to the eventual cellular uptake, intracellular trafficking, and expression or silencing of specific genes. Some of the more popular
nonviral nanodelivery systems, include polymeric micelles, dendrimers, lipid and polymeric nanoparticles, metal nanoparticles, quantum dots (QDs), and liposomes [1].
Kumaran Narayanan (ed.), Bio-Carrier Vectors: Methods and Protocols, Methods in Molecular Biology, vol. 2211,
https://doi.org/10.1007/978-1-0716-0943-9_4, © Springer Science+Business Media, LLC, part of Springer Nature 2021
43
Assessing Nucleic Acid: Cationic Nanoparticle Interaction
for Gene Delivery
Moganavelli Singh
Abstract
The assessment of the efficient binding between a nucleic acid and its associated nanoparticle is crucial for
gene delivery. Emerging from the extensive search for versatile gene carriers, are complexes formed between
nucleic acids and nonviral nanocarriers that promise to be viable alternatives to the predominantly viralbased gene delivery vehicles. However, much is still to be known about the exact structure and physicochemical properties of such nanocomplexes. This chapter will concentrate on cationic lipid, polymer, and
functionalized metal nanoparticles and their interaction with nucleic acids by direct conjugation or electrostatic interaction. Methods commonly employed to evaluate the nature and extent of nucleic acid interactions with cationic nanocarriers, such a nucleic acid binding, nuclease protection, and dye displacement
assays will be described. In addition, the ultrastructural morphology, size, and zeta potential of these
nanocomplexes, which are crucial for their cellular uptake and intracellular trafficking, will be assessed
using electron microscopy, fluorescent detection, and nanoparticle tracking analysis (NTA). These assays
have the ability to visualize and quantify the interaction and can also be used to complement each other, in
addition to providing confirmation of the formation of the relevant nanocomplexes.
Key words Nucleic acids, Nanoparticles, Nanocarriers, Interaction, Complexes, Gene delivery
1 Introduction
The exhaustive search for safe and versatile gene delivery systems
has produced an array of different systems to date, ranging from the
popular viral systems to lipids, polymers, and inorganic nanoparticles such as gold, selenium, silver, iron oxide, and platinum, to
name a few. Important to the use of these carrier systems are
complexes formed between the nucleic acid and the nanocarrier.
The exact structure and properties of such nanocomplexes is crucial
to the eventual cellular uptake, intracellular trafficking, and expression or silencing of specific genes. Some of the more popular
nonviral nanodelivery systems, include polymeric micelles, dendrimers, lipid and polymeric nanoparticles, metal nanoparticles, quantum dots (QDs), and liposomes [1].
Kumaran Narayanan (ed.), Bio-Carrier Vectors: Methods and Protocols, Methods in Molecular Biology, vol. 2211,
https://doi.org/10.1007/978-1-0716-0943-9_4, © Springer Science+Business Media, LLC, part of Springer Nature 2021
43