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Nat Protoc. 2007;2(12):3153-65.

Preferential delivery of the Sleeping Beauty transposon system to livers of mice by hydrodynamic injection.

Author information

1
Department of Genetics, Cell Biology and Development, Beckman Center for Transposon Research, Institute of Human Genetics, University of Minnesota, 6-160 Jackson Hall, 321 Church Street SE, Minneapolis, Minnesota 55455, USA.

Abstract

Nonviral, DNA-mediated gene transfer is an alternative to viral delivery systems for expressing new genes in cells and tissues. The Sleeping Beauty (SB) transposon system combines the advantages of viruses and naked DNA molecules for gene therapy purposes; however, efficacious delivery of DNA molecules to animal tissues can still be problematic. Here we describe the hydrodynamic delivery procedure for the SB transposon system that allows efficient delivery to the liver in the mouse. The procedure involves rapid, high-pressure injection of a DNA solution into the tail vein. The overall procedure takes <1 h although the delivery into one mouse requires only a few seconds. Successful injections result in expression of the transgene in 5-40% of hepatocytes 1 d after injection. Several weeks after injection, transgene expression stabilizes at approximately 1% of the level at 24 h, presumably owing to integration of the transposons into chromosomes.

PMID:
18079715
PMCID:
PMC2548418
DOI:
10.1038/nprot.2007.471
[Indexed for MEDLINE]
Free PMC Article

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