Development of genetic manipulation technology for excitatory and inhibitory control of the target neuron activity

在本研究中，我们的目的是开发一种新的技术，兴奋性和抑制性控制的活动的目标神经元类型。将抗人白细胞介素2受体α亚基的抗Tac单克隆抗体的单链可变区与Fc片段融合[anti-Tac（Fv）-Fc]与脂质体结合，制备免疫脂质体。将抗Tac抗体（Anti-Tac，Fv）与水泡性躯体炎病毒糖蛋白（VSV-G）的跨膜结构域连接，构建成抗Tac抗体（Anti-Tac，Fv）-VSV-G的杆状病毒表达载体。将这些细胞与脂质体融合以产生含有抗Tac（Fv）-VSV-G的蛋白脂质体。我们成功地减少免疫脂质体和蛋白脂质体的非特异性结合，通过添加未标记的对照脂质体到含有罗丹明标记的脂质体的溶液中。这一策略将有助于通过在免疫脂质体或蛋白脂质体中引入抑制离子通道的药物来控制神经元的活动。

In the present study, we aimed to develop a novel technology for excitatory and inhibitory control of the activity of target neuronal types. A single chain variable region of anti-Tac monoclonal antibody against human interleukin-2 receptor a-subunit was fused to Fc fragment[anti-Tac(Fv)-Fc] was bound to liposome to produce immunoliposome. Anti-Tac(Fv) was connected to the transmembrane domain of vesicular somatitis virus glycoprotein or VSV-G[anti-Tac(Fv)-VSV-G], and anti-Tac(Fv)-VSV-G was expressed in insect cells by the baculoviral vector system. These cells were fused to loposome to generate proteoliposome containing anti-Tac(Fv)-VSV-G. We succeeded in the reduction of nonspecific binding of immunoliposome and proteoliposome by adding unlabelled control liposome into the solution containing the rhodamine-labelled liposome. This strategy will be useful to control the activity of neurons by introducing some drugs that inhibit ion channels into immunoliposome or proteoliposome.