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Product lists for commonly used protocols from Cold Spring Harbor Protocols

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.

ABSTRACT

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-tansfer
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