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Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles

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Book cover Nanotechnology for Nucleic Acid Delivery

Part of the book series: Methods in Molecular Biology ((MIMB,volume 948))

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 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 poses significant challenges due to aggregation.

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Acknowledgements

We would like to thank Talar Tokatlian, Shiva Gojgini, Tim Martin, Sarah Plautz, and Tadas Kasputis for helpful comments and suggestions. We also like to thank the National Institutes of Health (R21EB009516, TS), the National Science Foundation (CAREER 0747539, TS), the American Heart Association (10SDG2640217), the University of Nebraska Foundation (Layman Funds), the Nebraska Research Initiative, NSF EPSCoR First Award, USDA CSREES-Nebraska (NEB-21-146), University of Nebraska-Lincoln IANR Strategic Investments Funds, and the J.A. Woollam Co. for financial support.

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Authors and Affiliations

  1. Department of Biological Systems Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA

    Angela K. Pannier

  2. Chemical and Biomolecular Engineering Department, University of California Los Angeles, Los Angeles, CA, USA

    Tatiana Segura

Authors
  1. Angela K. Pannier
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  2. Tatiana Segura
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Correspondence to Tatiana Segura .

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Editors and Affiliations

  1. , Department of Pharmacy, Ludwig-Maximilians-University Munich, Butenandstraße 5-13, Munich, 81377, Germany

    Manfred Ogris

  2. , Department of Pharmaceutical Sciences, Wayne State University, Mack Avenue 259, Detroit, 48201, USA

    David Oupicky

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Pannier, A.K., Segura, T. (2013). Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles. In: Ogris, M., Oupicky, D. (eds) Nanotechnology for Nucleic Acid Delivery. Methods in Molecular Biology, vol 948. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-140-0_11

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  • DOI: https://doi.org/10.1007/978-1-62703-140-0_11

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-139-4

  • Online ISBN: 978-1-62703-140-0

  • eBook Packages: Springer Protocols

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