1] Department of Biochemistry, University of Otago, Dunedin, New Zealand [2] Brain Health Research Centre, University of Otago, Dunedin, New Zealand.
Department of Biochemistry, University of Otago, Dunedin, New Zealand.
Gene Ther. 2015 May;22(5):357-64. doi: 10.1038/gt.2015.3. Epub 2015 Jan 29.
Lentiviral vectors have proved an effective method to deliver transgenes into the brain; however, they are often hampered by a lack of spread from the site of injection. Modifying the viral envelope with a portion of a rabies envelope glycoprotein can enhance spread in the brain by using long-range axon projections to facilitate retrograde transport. In this study, we generated two chimeric envelopes containing the extra-virion and transmembrane domain of rabies SADB19 or CVS-N2c with the intra-virion domain of vesicular stomatitis virus. Viral particles were packaged containing a green fluorescent protein reporter construct under the control of the phosphoglycerokinase promoter. Both vectors produced high-titer particles with successful integration of the glycoproteins into the particle envelope and significant transduction of neurons in vitro. Injection of the SADB19 chimeric viral vector into the lumbar spinal cord of adult mice mediated a strong preference for gene transfer to local neurons and axonal terminals, with retrograde transport to neurons in the brainstem, hypothalamus and cerebral cortex. Development of this vector provides a useful means to reliably target select populations of neurons by retrograde targeting.
慢病毒载体已被证明是将转基因递送到大脑中的有效方法；然而，它们常常受到注射部位传播范围有限的限制。通过使用长程轴突投射来促进逆行转运，用狂犬病包膜糖蛋白的一部分修饰病毒包膜，可以增强在大脑中的传播。在这项研究中，我们生成了两种嵌合包膜，包含狂犬病 SADB19 或 CVS-N2c 的额外病毒和跨膜结构域，以及水疱性口炎病毒的内部病毒结构域。病毒颗粒被包装，含有在磷酸甘油激酶启动子控制下的绿色荧光蛋白报告基因构建体。两种载体都产生了高滴度的颗粒，成功地将糖蛋白整合到颗粒包膜中，并显著转导了体外神经元。将 SADB19 嵌合病毒载体注射到成年小鼠的腰椎脊髓中，介导了对局部神经元和轴突末梢的强烈基因转移偏好，以及向脑干、下丘脑和大脑皮层神经元的逆行转运。这种载体的开发提供了一种通过逆行靶向可靠地靶向选择神经元群体的有用手段。
