Dreyer Jean-Luc
Division of Biochemistry, Department of Medicine, University of Fribourg, Fribourg, Switzerland.
Methods Mol Biol. 2010;614:3-35. doi: 10.1007/978-1-60761-533-0_1.
Lentiviral-mediated gene transfer in vivo or in cultured mammalian neurons can be used to address a wide variety of biological questions, to design animal models for specific neurodegenerative pathologies, or to test potential therapeutic approaches in a variety of brain disorders. Lentiviruses can infect nondividing cells, thereby allowing stable gene transfer in postmitotic cells such as mature neurons. An important contribution has been the use of inducible vectors: the same animal can thus be used repeatedly in the doxycycline-on or -off state, providing a powerful mean for assessing the function of a gene candidate in a disorder within a specific neuronal circuit. Furthermore, lentivirus vectors provide a unique tool to integrate siRNA expression constructs with the aim to locally knockdown expression of a specific gene, enabling to assess the function of a gene in a very specific neuronal pathway. Lentiviral vector-mediated delivery of short hairpin RNA results in persistent knockdown of gene expression in the brain. Therefore, the use of lentiviruses for stable expression of siRNA in brain is a powerful aid to probe gene functions in vivo and for gene therapy of diseases of the central nervous system. In this chapter, I review the applications of lentivirus-mediated gene transfer in the investigation of specific gene candidates involved in major brain disorders and neurodegenerative processes. Major applications have been in polyglutamine disorders, such as synucleinopathies and Parkinson's disease, or in investigating gene function in Huntington's disease, dystonia, or muscular dystrophy. Recently, lentivirus gene transfer has been an invaluable tool for evaluation of gene function in behavioral disorders such as drug addiction and attention-deficit hyperactivity disorder or in learning and cognition.
慢病毒介导的体内或培养的哺乳动物神经元基因转移可用于解决各种各样的生物学问题，设计特定神经退行性疾病的动物模型，或测试各种脑部疾病的潜在治疗方法。慢病毒可以感染非分裂细胞，从而允许在有丝分裂后细胞（如成熟神经元）中进行稳定的基因转移。一个重要的贡献是使用了诱导型载体：因此，同一动物可以在强力霉素开启或关闭状态下反复使用，这为评估特定神经元回路中候选基因在疾病中的功能提供了一个强大的手段。此外，慢病毒载体提供了一个独特的工具来整合siRNA表达构建体，目的是局部敲低特定基因的表达，从而能够在非常特定的神经元通路中评估基因的功能。慢病毒载体介导的短发夹RNA递送导致大脑中基因表达的持续敲低。因此，使用慢病毒在大脑中稳定表达siRNA是在体内探测基因功能和用于中枢神经系统疾病基因治疗的有力辅助手段。在本章中，我将回顾慢病毒介导的基因转移在研究参与主要脑部疾病和神经退行性过程的特定候选基因中的应用。主要应用于多聚谷氨酰胺疾病，如突触核蛋白病和帕金森病，或用于研究亨廷顿舞蹈病、肌张力障碍或肌肉萎缩症中的基因功能。最近，慢病毒基因转移已成为评估行为障碍（如药物成瘾和注意力缺陷多动障碍）或学习与认知中基因功能的宝贵工具。
