|
|
The following Cold Spring Harbor Laboratory Press protocols are brought to you by BioSupplyNet in a partnership with Sigma-Aldrich. A list of products is available to use in each protocol.
View the protocol on BioSupplyNet.com. |
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. |
Buy this book,
Z730033
|
| 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 | |
| Isopropyl alcohol | I9782 | Isopropyl alcohol, meets EP, BP, USP testing specifications | |
| 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 | |
| Polyvinyl alcohol (PVA) (m.w. 30,000-70,000; 87%-89% hydrolyzed) | 81369 | Poly(vinyl alcohol), standard, for GPC, 35,000 | |
| Tris-EDTA buffer (pH 8.0) | 93283 | Tris-EDTA buffer solution, BioUltra, for molecular biology, pH 8.0 | |
Back to Top
View all available Cold Spring Harbor Laboratory Press Protocols
Product Association Disclaimer: The Sigma-Aldrich products listed for this specific protocol were selected either to match or to supplement the products listed within the actual protocol. The products/reagents from Sigma-Aldrich have been qualified for usage, but may not have been validated for this specific application. Please refer to the detailed product description on the usage of specific products of interest.
|
