Generation of Adeno-Associated Virus Vector Enabling Functional Expression of Oxytocin Receptor and Fluorescence Marker Genes Using the Human eIF4G Internal Ribosome Entry Site Element
Generation of Adeno-Associated Virus Vector Enabling Functional Expression of Oxytocin Receptor and Fluorescence Marker Genes Using the Human eIF4G Internal Ribosome Entry Site Element
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DOI：
10.1271/bbb.90287
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发表时间：
2009-09-01
影响因子：
1.6
通讯作者：
Nishimori, Katsuhiko
中科院分区：
文献类型：
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作者：
Sato, Keisuke;Date, Shiori;Nishimori, Katsuhiko
我们开发了AAV-OXTR-IRES-Venus载体，功能上挽救了OXTR基因敲除小鼠脑内限制性区域的催产素受体(OXTR)基因。首先，我们选择了人eIF4G基因衍生的IRES与OXTR共表达荧光标记基因Venus。以筛选出的人eIF4G IRES为模板，构建了AAV-OXTR-IRES-Venus载体，向杏仁内侧核注射1亩L病毒液(9.4×10(7)VG)，可引起脑内Venus基因的表达。在转导该病毒载体的原代神经细胞中，用钙离子显像法检测OXTR的功能表达。
We developed the AAV-Oxtr-IRES-Venus vector to rescue the oxytocin receptor (Oxtr) gene functionally at restricted regions in the brains of Oxtr knockout mice. First we chose human eIF4G gene-derived IRES to co-express Venus, a fluorescent marker gene, with Oxtr. With selected human eIF4G IRES, we constructed the AAV-Oxtr-IRES-Venus vector, and it caused expression of the Venus gene in the brain when 1 mu l of viral solution (9.4 x 10(7) vg) was injected into the medial amygdaloid nucleus. In primary neuronal cells transduced with this viral vector and followed by oxytocin administration, functional expression of OXTR was detected by Ca2+ imaging assay.