Please cite as: CSH Protocols; 2008; doi:10.1101/pdb.prot4888

This Protocol
Right arrow Full Text
Right arrow Update/discuss this protocolDiscussion icon
Right arrow Alert me when this protocol is cited
Right arrow Alert me when comments are published
Right arrow Alert me if a correction is posted
Services
Right arrow Similar protocols in this database
Right arrow Alert me to new releases of protocols
Right arrow Save to Personal Folders
Right arrow Download to citation manager
Right arrow Printer-friendly versionPrinter-friendly version
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Vasir, J. K.
Right arrow Articles by Labhasetwar, V.
Right arrow Search for Related Content
PubMed
Right arrow Articles by Vasir, J. K.
Right arrow Articles by Labhasetwar, V.
Related Collections
Right arrow Molecular Biology, general
Right arrow Analysis of Gene Expression
Right arrow Expression of Cloned Genes
Right arrow Plasmids
Right arrow Vectors
Right arrow DNA Delivery/Gene Transfer
Right arrow DNA Delivery/Gene Transfer, general
Right arrow Non-Viral Methods
Right arrow Analysis of Gene Expression in Cultured Cells
Right arrow Cell Biology, general
Right arrow Cell Culture
Right arrow Preparation of Macromolecules and Introduction into Cells
Right arrowRelated Article
Right arrowRelated Protocols
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?
Legend icon

protocolProtocol

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.


INTRODUCTION

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.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?

Related Article

Protein Nanospheres for Gene Delivery
Sushma Kommareddy and Mansoor M. Amiji
CSH Protocols 2008: 30. [Abstract] [Full Text]

Related Protocols

Preparation and Loading of Gelatin Nanoparticles
Sushma Kommareddy and Mansoor M. Amiji
CSH Protocols 2008: 4885. [Abstract] [Full Text]

Intracellular Trafficking Studies Using Gold-Encapsulated Gelatin Nanoparticles
Sushma Kommareddy and Mansoor M. Amiji
CSH Protocols 2008: 4886. [Abstract] [Full Text]

Cell Transfection and Analysis Using DNA-Loaded Gelatin Nanoparticles
Sushma Kommareddy and Mansoor M. Amiji
CSH Protocols 2008: 4887. [Abstract] [Full Text]