Recombinant adeno-associated virus vector: use for transgene expression and anterograde tract tracing in the CNS
Recombinant adeno-associated virus vector: use for transgene expression and anterograde tract tracing in the CNS
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DOI：
10.1016/s0006-8993(98)00169-3
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发表时间：
1998-05-18
期刊：
影响因子：
2.9
通讯作者：
Saper, CB
中科院分区：
文献类型：
--
作者：
Chamberlin, NL;Du, B;Saper, CB
我们使用重组腺相关病毒载体（AAV）将外源基因绿色荧光蛋白（GFP）传递到体内成年大鼠中枢神经系统的成熟神经元中。小至 50 nl 的 AAV 显微注射可在注射部位转导数百个神经元。实际上没有逆行运输，因为每个大脑中发现远离表现出 GFP 免疫反应性的注射部位的神经元少于一个。基因产物 GFP 充满了整个神经元细胞质区室； GFP 免疫反应性在细胞体、轴突和神经末梢中很强。注射部位没有组织损伤，星形胶质细胞或小胶质细胞标记物的变化也表明没有致病性。根据白细胞侵袭判断没有炎症反应。转导细胞中的基因表达强劲且明显持久：感染后 12 周内没有表型逆转的证据。 AAV 是将外源基因引入成熟 CNS 神经元的优秀载体。它不仅可能是基因治疗的理想载体，而且含有 GFP 的 AAV 还提供了一种追踪中枢神经系统中长轴突通路的新策略，这对于当前的示踪剂（PHAL，生物素化葡聚糖）来说是困难的。 (C) 1998 Elsevier Science B.V.
We used a recombinant adeno-associated virus vector (AAV) to deliver a foreign gene, green fluorescent protein (GFP), into mature neurons in adult rat CNS in vivo. Microinjections of AAV as small as 50 nl transduced hundreds of neurons at the injection site. There was virtually no retrograde transport as fewer than one neuron per brain was found distant from the injection site that exhibited GFP immunoreactivity. The gene product, GFP, filled the entire neuronal cytoplasmic compartment; GFP immunoreactivity was robust in cell bodies, axons, and nerve terminals. There was no tissue damage at the injection sites or pathogenicity indicated by changes in astrocytic or microglial markers. There was no inflammatory response as judged by leukocytic invasion. Gene expression in transduced cells was robust and apparently permanent: there was no evidence of phenotypic reversion up to 12 weeks following infection. AAV is an excellent vector for introducing foreign genes into mature CNS neurons. Not only might it be an ideal vehicle for gene therapy, but also the GFP-containing AAV presents a new strategy for tracing long axonal pathways in the CNS, which is difficult with current tracers (PHAL, biotinylated dextrans). (C) 1998 Elsevier Science B.V.