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Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles.

Publication ,  Journal Article
Pannier, AK; Kozisek, T; Segura, T
Published in: Methods in molecular biology (Clifton, N.J.)
January 2019

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.

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Published In

Methods in molecular biology (Clifton, N.J.)

DOI

EISSN

1940-6029

ISSN

1064-3745

Publication Date

January 2019

Volume

1943

Start / End Page

177 / 197

Related Subject Headings

  • Transfection
  • Surface Properties
  • Plasmids
  • Nanoparticles
  • Mice
  • Hydrogels
  • Gene Expression Regulation
  • Developmental Biology
  • DNA
  • Cell Culture Techniques
 

Citation

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Pannier, A. K., Kozisek, T., & Segura, T. (2019). Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles. Methods in Molecular Biology (Clifton, N.J.), 1943, 177–197. https://doi.org/10.1007/978-1-4939-9092-4_12
Pannier, Angela K., Tyler Kozisek, and Tatiana Segura. “Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles.Methods in Molecular Biology (Clifton, N.J.) 1943 (January 2019): 177–97. https://doi.org/10.1007/978-1-4939-9092-4_12.
Pannier AK, Kozisek T, Segura T. Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles. Methods in molecular biology (Clifton, NJ). 2019 Jan;1943:177–97.
Pannier, Angela K., et al. “Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles.Methods in Molecular Biology (Clifton, N.J.), vol. 1943, Jan. 2019, pp. 177–97. Epmc, doi:10.1007/978-1-4939-9092-4_12.
Pannier AK, Kozisek T, Segura T. Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles. Methods in molecular biology (Clifton, NJ). 2019 Jan;1943:177–197.

Published In

Methods in molecular biology (Clifton, N.J.)

DOI

EISSN

1940-6029

ISSN

1064-3745

Publication Date

January 2019

Volume

1943

Start / End Page

177 / 197

Related Subject Headings

  • Transfection
  • Surface Properties
  • Plasmids
  • Nanoparticles
  • Mice
  • Hydrogels
  • Gene Expression Regulation
  • Developmental Biology
  • DNA
  • Cell Culture Techniques