Molecular Biology

Biodegradable Nanoparticles

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Preparation of Biodegradable Nanoparticles and Their Use in Transfection

Jaspreet K. Vasir and Vinod Labhasetwar
This protocol was adapted from "Biodegradable Nanoparticles," Chapter 56, in Gene Transfer: Delivery and Expression of DNA and RNA (eds. Friedmann and Rossi). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2007.

Biodegradable nanoparticles (NPs) are colloidal particles with a gene of interest encapsulated inside a polymeric matrix. They are typically 100 nm in diameter, and are formulated using FDA-approved, biodegradable, biocompatible polymers such as poly(D,L-lactide-co-glycolide) (PLGA) or polylactide (PLA). The NPs are taken up by cells via an endocytic process, and the encapsulated plasmid DNA entrapped in NPs is protected from degradation by both extra- and intracellular nucleases. It is released slowly, sustaining gene delivery and gene expression. In contrast, higher but transient gene expression is observed with lipid- or polymer-based complexes where most of the delivered DNA is available quickly for transfection. Thus, unlike other nonviral gene delivery systems, NPs thus constitute a sustained gene expression vector. Sustained gene expression is advantageous, especially when the half-life of the expressed protein is very low or when chronic gene delivery is required for therapeutic efficacy. This protocol describes a method for nanoencapsulation of DNA and the subsequent use of NPs for transfection.
Gene Transfer: Delivery and Expression of DNA and RNA Manual

Products Available for this Protocol
Protocol Material Description Product # Product Name Add to Cart
RPMI 1640 R8758 RPMI-1640 Medium, With L-glutamine and sodium bicarbonate, liquid, sterile-filtered, cell culture tested
Acetylated BSA (Ac-BSA) B2518 Albumin, Acetylated from bovine serum, lyophilized powder, for molecular biology
Triton X-100 T8787 Triton® X-100, for molecular biology
Chloroform C2432 Chloroform, ≥99%, contains amylenes as stabilizer
Luciferase Assay Substrate Luciferase Substrates  
PBS P5368 Phosphate buffered saline, pH 7.4, Biotechnology Performance Certified
Plasmid DNA D3404 Plasmid DNA from Escherichia coli RRI, pUC19, buffered aqueous solution
PLGA polymer (50:50 polylactide:glycolide ratio, inherent viscosity 1.32 dL/g in hexafluoroisopropanol) P2066 Poly(DL-lactide-co-glycolide), lactide:glycolide (65:35), mol wt 40,000-75,000

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