通过尾静脉高压注射将睡美人转座子系统优先递送至小鼠肝脏。

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

作者信息

Bell Jason B, Podetz-Pedersen Kelly M, Aronovich Elena L, Belur Lalitha R, McIvor R Scott, Hackett Perry B

机构信息

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.

出版信息

Nat Protoc. 2007;2(12):3153-65. doi: 10.1038/nprot.2007.471.

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.

非病毒、DNA介导的基因转移是在细胞和组织中表达新基因的病毒递送系统的替代方法。睡美人（SB）转座子系统结合了病毒和裸DNA分子在基因治疗方面的优势；然而，将DNA分子有效递送至动物组织仍然存在问题。在此，我们描述了用于SB转座子系统的流体动力学递送程序，该程序可实现向小鼠肝脏的高效递送。该程序包括将DNA溶液快速、高压注入尾静脉。整个程序耗时不到1小时，尽管向一只小鼠的递送仅需几秒钟。成功注射后，注射后1天，转基因在5%至40%的肝细胞中表达。注射后几周，转基因表达稳定在24小时时水平的约1%，这可能是由于转座子整合到染色体中所致。