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Gene Transfer to the Cerebellum

机译：基因转移到小脑

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摘要

There are several diseases for which gene transfer therapy to the cerebellum might be practicable. In these studies, we used recombinant Tag-deleted SV40-derived vectors (rSV40s) to study gene delivery targeting the cerebellum. These vectors transduce neurons and microglia very effectively in vitro and in vivo, and so we tested them to evaluate gene transfer to the cerebellum in vivo. Using a rSV40 vector carrying human immunodeficiency virus (HIV)-Nef with a C-terminal FLAG epitope, we characterized the distribution, duration, and cell types transduced. Rats received test and control vectors by stereotaxic injection into the cerebellum. Transgene expression was assessed 1, 2, and 4 weeks later by immunostaining of serial brain sections. FLAG epitope-expressing cells were seen, at all times after vector administration, principally detected in the Purkinje cells of the cerebellum, identified as immunopositive for calbindin. Occasional microglial cells were tranduced; transgene expression was not detected in astrocytes or oligodendrocytes. No inflammatory or other reaction was detected at any time. Thus, SV40-derived vectors can deliver effective, safe, and durable transgene expression to the cerebellum.
机译：有几种疾病可以通过基因转移疗法治疗到小脑。在这些研究中，我们使用重组标签缺失的SV40衍生载体（rSV40s）研究靶向小脑的基因传递。这些载体在体外和体内均非常有效地转导神经元和小胶质细胞，因此我们对其进行了测试，以评估体内基因向小脑的转移。使用带有人免疫缺陷病毒（HIV）-Nef和C端FLAG表位的rSV40载体，我们表征了转导的分布，持续时间和细胞类型。大鼠通过立体定向注射到小脑中接受测试和对照载体。 1、2和4周后通过连续脑切片的免疫染色评估转基因表达。在载体施用后的所有时间都观察到表达FLAG表位的细胞，主要在小脑的浦肯野细胞中检测到，被鉴定为钙结合蛋白的免疫阳性。偶尔转导小胶质细胞；在星形胶质细胞或少突胶质细胞中未检测到转基因表达。任何时候均未检测到炎症或其他反应。因此，源自SV40的载体可以将有效，安全和持久的转基因表达传递至小脑。

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  • 来源
    《The Cerebellum》 |2010年第4期|587-597|共11页
  • 作者

    Jean-Pierre Louboutin; Beverly A. S. Reyes; Elisabeth J. Van Bockstaele; David S. Strayer;

  • 作者单位

    Department of Pathology Jefferson Medical College Thomas Jefferson University 1020 Locust Street Room 255 Philadelphia PA 19107 USA;

    Department of Neurosurgery Farber Institute for Neurosciences Thomas Jefferson University Philadelphia PA 19107 USA;

    Department of Neurosurgery Farber Institute for Neurosciences Thomas Jefferson University Philadelphia PA 19107 USA;

    Department of Pathology Jefferson Medical College Thomas Jefferson University 1020 Locust Street Room 255 Philadelphia PA 19107 USA;

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  • 原文格式 PDF
  • 正文语种 eng
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  • 关键词

    SV40; Gene transfer; Cerebellum;

    机译：SV40;基因转移;小脑;

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