Electroporation enhances reporter gene expression following delivery of naked plasmid DNA to the lung

Pringle, Ian A; McLachlan, Gerry; Collie, David D S; Sumner-Jones, Stephanie G; Lawton, Anna E; Tennant, Peter; Baker, Alison; Gordon, Catherine; Blundell, Richard; Varathalingam, Anusha; Davies, Lee A; Schmid, Ralph A; Cheng, Seng H; Porteous, David J; Gill, Deborah R; Hyde, Stephen C (2007). Electroporation enhances reporter gene expression following delivery of naked plasmid DNA to the lung. Journal of gene medicine, 9(5), pp. 369-80. Chichester: Wiley 10.1002/jgm.1026

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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.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Gastro-intestinal, Liver and Lung Disorders (DMLL) > Clinic of Thoracic Surgery

UniBE Contributor:

Schmid, Ralph

ISSN:

1099-498X

ISBN:

17410613

Publisher:

Wiley

Language:

English

Submitter:

Factscience Import

Date Deposited:

04 Oct 2013 15:01

Last Modified:

04 May 2014 23:18

Publisher DOI:

10.1002/jgm.1026

PubMed ID:

17410613

Web of Science ID:

000247051100006

URI:

https://boris.unibe.ch/id/eprint/26394 (FactScience: 70115)

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