Document Detail


Electroporation enhances reporter gene expression following delivery of naked plasmid DNA to the lung.
MedLine Citation:
PMID:  17410613     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Existing methods of non-viral airway gene transfer suffer from low levels of efficiency. Electroporation has been used to enhance gene transfer in a range of tissues. Here we assess the usefulness of electroporation for enhancing gene transfer in the lungs of mice and sheep. METHODS: Naked plasmid DNA (pDNA) expressing either luciferase or green fluorescent protein (GFP) was delivered to mouse lungs by instillation. Following surgical visualisation, the lungs were directly electroporated and the level and duration of luciferase activity was assessed and cell types that were positive for GFP were identified in lung cryosections. Naked pDNA was nebulised to the sheep lung and electrodes attached to the tip of a bronchoscope were used to electroporate airway segment bifurcations, Luciferase activity was assessed in electroporated and control non-electroporated regions, after 24 h. RESULTS: Following delivery of naked pDNA to the mouse lung, electroporation resulted in up to 400-fold higher luciferase activity than naked pDNA alone when luciferase was under the control of a cytomegalovirus (CMV) promoter. Following delivery of a plasmid containing the human polyubiquitin C (UbC) promoter, electroporation resulted in elevated luciferase activity for at least 28 days. Visualisation of GFP indicated that electroporation resulted in increased GFP detection compared with non-electroporated controls. In the sheep lung electroporation of defined sites in the airways resulted in luciferase activity 100-fold greater than naked pDNA alone. CONCLUSIONS: These results indicate that electroporation can be used to enhance gene transfer in the lungs of mice and sheep without compromising the duration of expression.
Authors:
Ian A Pringle; Gerry McLachlan; David D S Collie; Stephanie G Sumner-Jones; Anna E Lawton; Peter Tennant; Alison Baker; Catherine Gordon; Richard Blundell; Anusha Varathalingam; Lee A Davies; Ralph A Schmid; Seng H Cheng; David J Porteous; Deborah R Gill; Stephen C Hyde
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The journal of gene medicine     Volume:  9     ISSN:  1099-498X     ISO Abbreviation:  J Gene Med     Publication Date:  2007 May 
Date Detail:
Created Date:  2007-05-07     Completed Date:  2007-08-30     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  9815764     Medline TA:  J Gene Med     Country:  England    
Other Details:
Languages:  eng     Pagination:  369-80     Citation Subset:  IM    
Copyright Information:
Copyright (c) 2007 John Wiley & Sons, Ltd.
Affiliation:
GeneMedicine Research Group, Nuffield Department of Clinical Laboratory Sciences, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, UK.
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MeSH Terms
Descriptor/Qualifier:
Animals
Electroporation*
Gene Expression
Gene Transfer Techniques*
Genes, Reporter / genetics*
Green Fluorescent Proteins / genetics
Humans
Kinetics
Luciferases / genetics
Lung / cytology*,  metabolism
Mice
Plasmids / genetics*
Promoter Regions, Genetic
Sheep
Chemical
Reg. No./Substance:
147336-22-9/Green Fluorescent Proteins; EC 1.13.12.-/Luciferases

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


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