Deletion of the Virion Host Shut-off Gene Enhances Neuronal-Selective Transgene Expression from an HSV Vector Lacking Functional IE Genes.
Deletion of the Virion Host Shut-off Gene Enhances Neuronal-Selective Transgene Expression from an HSV Vector Lacking Functional IE Genes.
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DOI：
10.1016/j.omtm.2017.06.001
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发表时间：
2017-09-15
期刊：
影响因子：
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通讯作者：
Glorioso JC
中科院分区：
文献类型：
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作者：
Miyagawa Y;Verlengia G;Reinhart B;Han F;Uchida H;Zucchini S;Goins WF;Simonato M;Cohen JB;Glorioso JC
单纯疱疹病毒（HSV）在神经元中建立终身潜伏期的能力表明，HSV衍生的载体有望将基因递送到神经系统。然而，载体毒性和转基因沉默已经对将载体应用于脑造成了显著障碍。最近，我们描述了一种载体缺陷的所有立即早期基因表达和删除的两个独特的基因组片段之间的联合区域，证明能够扩展在非神经元细胞中的转基因表达。持续表达需要接近潜伏期位点的边界元素。如本文所证实的，我们还发现引入ICP4基因座的转基因盒在神经元中高度活跃，但在原代成纤维细胞中沉默。值得注意的是，我们观察到去除病毒体宿主关闭（vhs）基因进一步提高了神经元中的转基因表达，而不诱导病毒基因的表达。在大鼠海马体中，vhs缺失的载体在至少1个月的时间内仅在神经元中表现出强有力的转基因表达，而没有毒性或炎症的证据。这种HSV载体设计有望将基因递送到大脑，包括持久表达大型或复杂的转基因盒。
The ability of herpes simplex virus (HSV) to establish lifelong latency in neurons suggests that HSV-derived vectors hold promise for gene delivery to the nervous system. However, vector toxicity and transgene silencing have created significant barriers to vector applications to the brain. Recently, we described a vector defective for all immediate-early gene expression and deleted for the joint region between the two unique genome segments that proved capable of extended transgene expression in non-neuronal cells. Sustained expression required the proximity of boundary elements from the latency locus. As confirmed here, we have also found that a transgene cassette introduced into the ICP4 locus is highly active in neurons but silent in primary fibroblasts. Remarkably, we observed that removal of the virion host shutoff (vhs) gene further improved transgene expression in neurons without inducing expression of viral genes. In rat hippocampus, the vhs-deleted vector showed robust transgene expression exclusively in neurons for at least 1 month without evidence of toxicity or inflammation. This HSV vector design holds promise for gene delivery to the brain, including durable expression of large or complex transgene cassettes.