Document Detail


Recent advances in magnetofection and its potential to deliver siRNAs in vitro.
MedLine Citation:
PMID:  19301645     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This chapter describes how to design and conduct experiments to deliver siRNA to adherent mammalian cells in vitro by magnetic force-assisted transfection using self-assembled complexes of small interfering RNA (siRNA) and cationic lipids or polymers that are associated with magnetic nanoparticles. These magnetic complexes are targeted to the cell surface by the application of a magnetic gradient field. In this chapter, first we describe the synthesis of magnetic nanoparticles for magnetofection and the association of siRNA with the magnetic components of the transfection complex. Second, a simple protocol is described in order to evaluate magnetic responsiveness of the magnetic siRNA transfection complexes and estimate the complex loading with magnetic nanoparticles. Third, protocols are provided for the preparation of magnetic lipoplexes and polyplexes of siRNA, magnetofection, downregulation of gene expression, and the determination of cell viability. The addition of INF-7 peptide, a fusogenic peptide, to the magnetic transfection triplexes improved gene silencing in HeLa cells. The described protocols are also valuable for screening vector compositions and novel magnetic nanoparticle preparations to optimize siRNA transfection by magnetofection in every cell type.
Authors:
Olga Mykhaylyk; Olivier Zelphati; Edelburga Hammerschmid; Martina Anton; Joseph Rosenecker; Christian Plank
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  487     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-03-23     Completed Date:  2009-04-16     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  111-46     Citation Subset:  IM    
Affiliation:
Institute of Experimental Oncology, Technische Universität München, Munich, Germany.
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MeSH Terms
Descriptor/Qualifier:
Carcinoma, Papillary / metabolism,  therapy
Cell Line, Tumor
Cell Proliferation
Gene Silencing
Gene Therapy / methods
Gene Transfer Techniques*
Genetic Vectors
Green Fluorescent Proteins / administration & dosage,  metabolism
Hela Cells
Humans
Lung Neoplasms / metabolism,  therapy
Magnetics*
Nanoparticles / chemistry
Peptides / antagonists & inhibitors,  genetics,  metabolism*
RNA, Small Interfering / administration & dosage*,  genetics
Transfection / methods*
Chemical
Reg. No./Substance:
0/INF7 peptide; 0/Peptides; 0/RNA, Small Interfering; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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