Protocol abstract
Nature Protocols 2, - 3146 - 3152 (2007)
Published online: 6 December 2007 | doi:10.1038/nprot.2007.460
Subject Categories: Genetic modification | Model organisms
Lung-directed gene therapy in mice using the nonviral Sleeping Beauty transposon system
Lalitha R Belur1, Kelly Podetz-Pedersen1, Joel Frandsen1 & R Scott McIvor1
Abstract
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.
- The Arnold and Mabel Beckman Center for Transposon Research, Gene Therapy Program, Institute of Human Genetics, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Correspondence to: R Scott McIvor1 e-mail: mcivor@mail.med.umn.edu
Correspondence to: Lalitha R Belur1 e-mail: belur001@umn.edu
nature-products
A-Z product listing
- Acepromazine maleate(Phoenix Pharmaceutical Incorporated)
- Balb/c mice(Jackson labs)
- Butorphanol tartrate(Fort Dodge Animal Health)
- D-Luciferin firefly luciferase substrate, potassium salt(Xenogen Corporation/Caliper Life Sciences)
- Eight-week-old C57BL/6 mice(National Cancer Institute)
- Endo-Free plasmid maxi kit(Qiagen)
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