Ehrengruber Markus U, Schlesinger Sondra, Lundstrom Kenneth
Department of Biology, Kantonsschule Hohe Promenade, Zurich, Switzerland.
Curr Protoc Neurosci. 2011 Oct;Chapter 4:Unit 4.22. doi: 10.1002/0471142301.ns0422s57.
Alphaviral vectors based on Semliki Forest virus and Sindbis virus infect many host cell types, causing rapid and high-level transgene expression. In the CNS, Semliki Forest virus and Sindbis virus exhibit an outstanding preference for neurons rather than glial cells, compared to other viruses. Generation of high-titer virus stocks is rapid (less than two days) and typically requires biosafety level 1 or 2 containment. Wild-type vectors are cytotoxic, permitting short-term transgene expression. However, mutant vectors with decreased cytotoxicity, to prolong host cell survival, have been developed. They also increase transgene expression and cellular co-infection, permitting heteromeric protein expression in individual cells. In addition, mutants with temperature-dependent control of transgene expression and altered host cell preference to target interneurons and astrocytes rather than principal neurons are available. Other alphavirus vectors based on Venezuelan equine encephalitis and Eastern equine encephalitis virus replicons have been engineered, too. Alphavirus vectors have been successfully used not only in neuroscience, but also for other applications including drug discovery, structural biology, vaccine development, and cancer therapy.
基于辛德毕斯病毒和塞姆利基森林病毒的甲病毒载体可感染多种宿主细胞类型，实现快速且高水平的转基因表达。在中枢神经系统中，相较于其他病毒，塞姆利基森林病毒和辛德毕斯病毒对神经元而非神经胶质细胞表现出显著的偏好。高滴度病毒储备的产生速度很快（不到两天），通常需要生物安全1级或2级防护。野生型载体具有细胞毒性，只能实现短期转基因表达。然而，为了延长宿主细胞存活时间，人们已开发出细胞毒性降低的突变载体。它们还能增加转基因表达和细胞共感染，从而在单个细胞中实现异源蛋白表达。此外，还有一些突变体，其转基因表达受温度调控，宿主细胞偏好发生改变，可靶向中间神经元和星形胶质细胞而非主要神经元。基于委内瑞拉马脑炎病毒和东部马脑炎病毒复制子的其他甲病毒载体也已构建成功。甲病毒载体不仅在神经科学领域得到了成功应用，还用于包括药物发现、结构生物学、疫苗开发和癌症治疗在内的其他应用。
