Peterson Elise B, Mastrangelo Michael A, Federoff Howard J, Bowers William J
Center for Aging and Developmental Biology, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
Mol Ther. 2007 Oct;15(10):1848-55. doi: 10.1038/sj.mt.6300267. Epub 2007 Jul 24.
A novel bipartite vector system consisting of the herpes simplex virus (HSV) amplicon and the Sleeping Beauty(SB) transposon was previously shown to efficiently deliver a "transgenon" (integrating transgene) in utero. This vector platform facilitated long-term transgenon expression specifically within neurons and neuronal precursor cells of the rodent brain. However, the mechanism underlying the neurospecificity of the HSV/SB amplicon in the setting of mouse embryogenesis is unknown. We find that embryonic cells expressing the Sox1 "neurocompetence" transcription factor represent the primary targets for HSV amplicon transduction in utero. These cells, which comprise the ependymal and subventricular zones (SVZs), express significant levels of high-mobility-group protein B1 (HMGB1), a co-factor shown to facilitate SB-mediated transposition. Using a conventional, non-integrating amplicon expressing Cre recombinase to "tag" transduced cells embryonically in ROSA26 Cre indicator mice in utero, we found transduced cells were exclusively of the neuronal lineage but that in comparison to HSV/SB-mediated in utero delivery, staining patterns were less widespread and "tagged" neuroprogenitor cells were absent. Our findings demonstrate that in utero HSV/SB amplicon gene transfer is primarily neurospecific owing to viral tropism and target cell populations present embryonically, where multi-potent cells of the developing embryo are supportive of SB-driven transposition.
一种由单纯疱疹病毒（HSV）扩增子和睡美人（SB）转座子组成的新型双组分载体系统，先前已证明其能在子宫内有效递送“转基因子”（整合转基因）。该载体平台促进了转基因子在啮齿动物大脑的神经元和神经前体细胞内的长期表达。然而，在小鼠胚胎发育过程中，HSV/SB扩增子神经特异性的潜在机制尚不清楚。我们发现，表达Sox1“神经能力”转录因子的胚胎细胞是子宫内HSV扩增子转导的主要靶标。这些细胞包括室管膜和脑室下区（SVZ），表达高水平的高迁移率族蛋白B1（HMGB1），这是一种已证明能促进SB介导转座的辅助因子。利用一种表达Cre重组酶的传统非整合扩增子，在子宫内对ROSA26 Cre指示小鼠胚胎中的转导细胞进行“标记”，我们发现转导细胞仅为神经谱系，但与HSV/SB介导的子宫内递送相比，染色模式分布较窄且不存在“标记”的神经祖细胞。我们的研究结果表明，子宫内HSV/SB扩增子基因转移主要具有神经特异性，这是由于病毒嗜性和胚胎期存在的靶细胞群体所致，发育胚胎中的多能细胞支持SB驱动的转座。
