Surface- and hydrogel-mediated delivery of nucleic acid nanoparticles

Angela K. Pannier, Tyler Kozisek, Tatiana Segura

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Gene expression within a cell population can be directly altered through gene delivery approaches. Traditionally for nonviral delivery, plasmids or siRNA molecules, encoding or targeting the gene of interest, are packaged within nanoparticles. These nanoparticles are then delivered to the media surrounding cells seeded onto tissue culture plastic; this technique is termed bolus delivery. Although bolus delivery is widely utilized to screen for efficient delivery vehicles and to study gene function in vitro, this delivery strategy may not result in efficient gene transfer for all cell types or may not identify those delivery vehicles that will be efficient in vivo. Furthermore, bolus delivery cannot be used in applications where patterning of gene expression is needed. In this chapter, we describe methods that incorporate material surfaces (i.e., surface-mediated delivery) or hydrogel scaffolds (i.e., hydrogel-mediated delivery) to efficiently deliver genes. This chapter includes protocols for surface-mediated DNA delivery focusing on the simplest and most effective methods, which include nonspecific immobilization of DNA complexes (both polymer and lipid vectors) onto serum-coated cell culture polystyrene and self-assembled monolayers (SAMs) of alkanethiols on gold. Also, protocols for the encapsulation of DNA/cationic polymer nanoparticles into hydrogel scaffolds are described, including methods for the encapsulation of low amounts of DNA (<0.2 μg/μl) and high amounts of DNA (>0.2 μg/μl) since incorporation of high amounts of DNA pose significant challenges due to aggregation.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages177-197
Number of pages21
DOIs
Publication statusPublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1943
ISSN (Print)1064-3745

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Keywords

  • Gene delivery
  • Hydrogel
  • Nonviral
  • Surface-mediated
  • Transfection

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Pannier, A. K., Kozisek, T., & Segura, T. (2019). Surface- and hydrogel-mediated delivery of nucleic acid nanoparticles. In Methods in Molecular Biology (pp. 177-197). (Methods in Molecular Biology; Vol. 1943). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9092-4_12