Huang Z Josh, Taniguchi Hiroki, He Miao, Kuhlman Sandra
Cold Spring Harb Protoc. 2014 Feb 1;2014(2):190-4. doi: 10.1101/pdb.prot080382.
Virus-mediated gene delivery is a powerful strategy for labeling and manipulating neurons in mammalian brains. A major drawback of this gene delivery method has been the lack of cell-type specificity. However, methods that combine Cre-knockin mice and Cre-activated adeno-associated virus (AAV) have now been developed to achieve high-level, stable, and cell-type-specific gene expression. Here, we describe a protocol for the design, production, and delivery of Cre-dependent AAVs. This method is simple and highly efficient, and allows chronic live imaging of defined classes of synapses in the mouse brain.
病毒介导的基因传递是标记和操纵哺乳动物大脑中神经元的一种强大策略。这种基因传递方法的一个主要缺点是缺乏细胞类型特异性。然而,现在已经开发出将Cre敲入小鼠和Cre激活的腺相关病毒(AAV)相结合的方法,以实现高水平、稳定和细胞类型特异性的基因表达。在这里,我们描述了一种用于设计、生产和递送依赖Cre的AAV的方案。该方法简单且高效,能够对小鼠大脑中特定类型的突触进行长期活体成像。