Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles

  • Angela K. Pannier
  • Tyler Kozisek
  • Tatiana SeguraEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1943)


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.

Key words

Gene delivery Hydrogel Surface-mediated Transfection Nonviral 



We would like to thank Andrew Hamann for unpublished results. We also like to thank the National Institutes of Health (R21EB009516, T.S.; 1 DP2 EB025760-01, A.K.P.), the National Science Foundation (CAREER 0747539, T.S.; CAREER CBET-1254415, A.K.P.) the American Heart Association (10SDG2640217), and USDA CSREES-Nebraska (NEB-21-146).


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Angela K. Pannier
    • 1
  • Tyler Kozisek
    • 1
  • Tatiana Segura
    • 2
    • 3
    Email author
  1. 1.Department of Biological Systems EngineeringUniversity of Nebraska-LincolnLincolnUSA
  2. 2.Department of Biomedical EngineeringDuke UniversityDurhamUSA
  3. 3.Neurology and DermatologyDuke University School of MedicineDurhamUSA

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