Document Detail


Biolistic Transfection of Human Embryonic Kidney (HEK) 293 Cells.
MedLine Citation:
PMID:  23104338     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Due to its excellent transfectability, the human embryonic kidney (HEK) 293 cell line is widely used as an in vitro model system for transfection experiments. Particle bombardment, or biolistics technology, provides a physical transfection approach that can deliver transgene materials efficiently into many different cell lines. Transfection of 293 cells by gene gun, allows examination of transgene expression in epithelial cells, as well as studies concerning a variety of questions in neurobiology. The present study of transfection of HEK 293 cells by biolistics technology uses the plasmids gWIZ-luc encoding luciferase and gWIZ-GFP encoding green fluorescence protein (GFP) as model transgenes. This system can be routinely used at varying bombarding conditions that can be adjusted according to experimental requirements and purpose, such as gene gun helium pressure, the sizes and the amount of the gold particles and the length of the spacer. The results obtained show that the Bio-Rad spacer for the gene gun should be optimized for travel distance and spreading of gold particles over a relatively small area, when used for biolistic transfection of cells dispersed in multi-well plate.
Authors:
Xiongwei Li; Masaki Uchida; H Oya Alpar; Peter Mertens
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  940     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-10-29     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  119-32     Citation Subset:  IM    
Affiliation:
Department of Pharmaceutics, School of Pharmacy, University of London, London, UK.
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