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首页> 外文期刊>Gene Therapy and Molecular Biology >The pathway of uptake of SV40 pseudovirions packaged in vitro: from MHC class I receptors to thenucleus
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The pathway of uptake of SV40 pseudovirions packaged in vitro: from MHC class I receptors to thenucleus

机译：体外包装的SV40假病毒颗粒的吸收途径：从I类MHC受体到细胞核

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摘要

SV40 vectors packaged in vitro are an efficient delivery system in vitro and in vivo using plasmids up to 17.7 kb, with or without SV40 sequences. Using confocal microscopy, we followed the pathway of SV40 pseudovirions in human lymphoblastoid cells, which are rich in MHC I receptors, using fluorescence-tagged DNA and an antibody against the main capsid protein, VP1. The wild-type SV40 virus as well as the pseudovirions enter the cells after binding to MHC I. However, the MHC I route is not the only way that SV40 pseudovirions enter cells. From the cell surface, the vectors progress through the Golgi to the ER, where they are unpackaged. Only the reporter DNA proceeds to the nucleus; VP1 remains at the ER. Results indicate that some of the reporter DNA, carried by these vectors, is trapped in the ER. Delivery of DNA plasmids which harbor nuclear localization sequences, such as the enhancer of wild-type SV40 or the cPPT sequence from the HIV-1 virus upstream from the GFP cDNA, did not improve GFP expression. However, improved expression from the EGFP reporter gene carried by SV40 vectors was achieved using the histone deacetylase inhibitor, TSA.
机译：体外包装的SV40载体是一种有效的体外和体内递送系统，使用高达17.7 kb的质粒（带有或不带有SV40序列）。使用共聚焦显微镜，我们使用荧光标记的DNA和针对主要衣壳蛋白VP1的抗体，在富含MHC I受体的人淋巴母细胞中追踪SV40假病毒颗粒的途径。野生型SV40病毒以及假病毒颗粒与MHC I结合后进入细胞。但是，MHC I途径并不是SV40假病毒颗粒进入细胞的唯一途径。载体从细胞表面穿过高尔基体到达内质网，在这里它们没有包装。只有报告DNA进入核。 VP1保留在ER。结果表明，这些载体携带的一些报道基因DNA被捕获在ER中。携带核定位序列的DNA质粒，例如野生型SV40的增强子或cPPT序列，是从GFP cDNA上游的HIV-1病毒中转移而来，并没有改善GFP的表达。但是，使用组蛋白脱乙酰基酶抑制剂TSA可实现SV40载体携带的EGFP报告基因表达的改善。

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