Belur Lalitha R, Podetz-Pedersen Kelly, Frandsen Joel, McIvor R Scott
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.
Nat Protoc. 2007;2(12):3146-52. doi: 10.1038/nprot.2007.460.
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.
睡美人（SB）转座子是一种整合性非病毒质粒系统。在此，我们描述了一种使用DNA/聚乙烯亚胺（PEI）复合物通过SB介导进行转基因递送以在小鼠肺部实现长期表达的方案。该方案可用于将任何基于质粒的载体系统递送至小鼠肺部，不过只有在使用SB转座子或其他整合载体系统时才能获得长期转基因表达。本方案的步骤包括制备DNA-PEI复合物以及将复合物注射到小鼠的侧尾静脉中。我们还提供了使用体内生物发光成像和肺匀浆酶促测定来评估转基因表达的方案。从制备DNA-PEI复合物到分析瞬时转基因表达，整个过程可在24小时内完成。
