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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Delivery and long-term expression of a 135 kb LDLR genomic DNA locus in vivo by hydrodynamic tail vein injection
Journal of Gene Medicine, Volume 9, No. 6, Year 2007
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Description
Background: The delivery of a complete genomic DNA locus in vivo may prove advantageous for complementation gene therapy, especially when physiological regulation of gene expression is desirable. Hydrodynamic tail vein injection has been shown to be a highly efficient means of non-viral delivery of plasmid DNA to the liver. Here, we apply hydrodynamic tail vein injection to deliver and express large genomic DNA inserts >100 kb in vivo. Methods: Firstly, a size series (12-172 kb) of bacterial artificial chromosome (BAC) plasmids, carrying human genomic DNA inserts, episomal retention elements, and the enhanced green fluorescent protein (EGFP) reporter gene, was delivered to mice by hydrodynamic tail vein injection. Secondly, an episomal BAC vector carrying the whole genomic DNA locus of the human low-density lipoprotein receptor (LDLR) gene, and an expression cassette for the LacZ reporter gene, was delivered by the same method. Results: We show that the efficiency of delivery is independent of vector size, when an equal number of plasmid molecules are used. We also show, by LacZ reporter gene analysis, that BAC delivery within the liver is widespread. Finally, BAC-end PCR, RT-PCR and immunohistochemistry demonstrate plasmid retention and long-term expression (4 months) of human LDLR in transfected hepatocytes. Conclusion: This is the first demonstration of somatic delivery and long-term expression of a genomic DNA transgene >100 kb in vivo and shows that hydrodynamic tail vein injection can be used to deliver and express large genomic DNA transgenes in the liver. Copyright ©2007 John Wiley & Sons, Ltd.
Authors & Co-Authors
Waddington, Simon N.
United Kingdom, London
Imperial College London
United Kingdom, London
Ucl Medical School
Coutelle, Charles C.
United Kingdom, London
Imperial College London
Channon, Keith Michael
United Kingdom, Oxford
University of Oxford
Wade-Martins, Richard
United Kingdom, Oxford
University of Oxford
Statistics
Citations: 40
Authors: 4
Affiliations: 3
Identifiers
Doi:
10.1002/jgm.1041
ISSN:
1099498X
Research Areas
Genetics And Genomics