Protocol abstract
Nature Protocols 2, - 3153 - 3165 (2007)
Published online: 6 December 2007 | doi:10.1038/nprot.2007.471
Subject Categories: Genetic modification | Model organisms
Preferential delivery of the Sleeping Beauty transposon system to livers of mice by hydrodynamic injection
Jason B Bell1, Kelly M Podetz-Pedersen1, Elena L Aronovich1, Lalitha R Belur1, R Scott McIvor1,2 & Perry B Hackett1
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 
1% of the level at 24 h, presumably owing to integration of the transposons into chromosomes.
- 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.
- Discovery Genomics, Inc., 614 McKinley Place NE, Minneapolis, Minnesota 55413, USA.
Correspondence to: Perry B Hackett1 e-mail: perryh@umn.edu
nature-products
A-Z product listing
- 10-ml sterile vacutainer tubes(Becton Dickenson)
- acepromazine maleate(Phoenix Pharmaceutical Incorporated)
- butorphanol tartrate(Fort Dodge Animal Health)
- D-Luciferin firefly luciferase substrate, potassium salt(Xenogen Corporation)
- DNeasy Tissue Kit(Qiagen)
- Dulbecco's PBS(Cambrix Bio Science)
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