Unité Neurobiologie Intégrative des Systèmes Cholinergiques, CNRS URA 2182, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.
FASEB J. 2010 Mar;24(3):723-30. doi: 10.1096/fj.09-139790. Epub 2009 Oct 26.
Lentiviral expression vectors are powerful tools for gene therapy and long-term gene expression/repression in the mammalian brain. However, no specificity of transduction has been reported so far in the central nervous system. Here we have developed a novel system to achieve a neuronal subtype specific expression in either dopaminergic (DA) or GABAergic neurons. We employed a delivery strategy by which the transgene is not expressed until its activation by Cre recombinase. We successfully tested the system in vitro and then used this novel lentivector, containing loxP sites, in 2 different transgenic mouse lines expressing Cre either in DA or in GABAergic neurons. In both lines the reporter gene was detected exclusively in Cre-positive cells, demonstrating that with this experimental approach we were able to achieve completely specific expression of transgenes delivered by lentiviral vectors. This universal system can be applied to all neural subtypes making use of the growing number of specific Cre driver lines.- Tolu, S., Avale, M. E., Nakatani, H., Pons, S., Parnaudeau, S., Tronche, F., Vogt, A., Monyer, H., Vogel, R., de Chaumont, F., Olivo-Marin, J.-C., Changeux, J.-P., Maskos, U. A versatile system for the neuronal subtype specific expression of lentiviral vectors.
慢病毒表达载体是基因治疗和哺乳动物大脑中长期基因表达/抑制的有力工具。然而，迄今为止，在中枢神经系统中还没有报道过转导的特异性。在这里，我们开发了一种新的系统，可以在多巴胺能 (DA) 或 GABA 能神经元中实现神经元亚型特异性表达。我们采用了一种传递策略，其中转基因在 Cre 重组酶激活之前不表达。我们在体外成功测试了该系统，然后在 2 种不同的转基因小鼠品系中使用了这种新型包含 loxP 位点的慢病毒载体，该品系在 DA 或 GABA 能神经元中表达 Cre。在这两种品系中，报告基因仅在 Cre 阳性细胞中检测到，表明通过这种实验方法，我们能够实现慢病毒载体传递的转基因的完全特异性表达。该通用系统可应用于所有神经亚型，利用越来越多的特异性 Cre 驱动线。
