Lung-directed gene therapy in mice using the nonviral Sleeping Beauty transposon system.
The Sleeping Beauty (SB) transposon is an integrative nonviral plasmid system. Here, we describe a protocol for SB-mediated transgene delivery using DNA/polyethyleneimine (PEI) complexes for long-term expression in mouse lungs. This protocol can be used for delivery of any plasmid-based vector system to mouse lungs, although long-term transgene expression will be obtained only when using the SB transposon or other integrating vector systems. The stages of this protocol are preparation of DNA-PEI complexes and injection of the complexes into the lateral tail vein of mice. We also provide protocols for assessing transgene expression using in vivo bioluminescence imaging and enzymatic assay of lung homogenates. The procedure can be completed within 24 h, starting from preparation of DNA-PEI complexes to analysis of transient transgene expression.
Belur
Lalitha R
LR
The Arnold and Mabel Beckman Center for Transposon Research, Institute of Human Genetics, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455, USA. belur001@umn.edu
Podetz-Pedersen
Kelly
K
Frandsen
Joel
J
McIvor
R Scott
RS
D000818:Animals; D004251:DNA Transposable Elements / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D008168:Lung / Q000378:metabolism*; D051379:Mice; D009928:Organ Specificity; D010957:Plasmids / Q000235:genetics; D019076:Transgenes / Q000235:genetics
D004251:DNA Transposable Elements