A Guide to Creating and Testing New INTRSECT Constructs.
A Guide to Creating and Testing New INTRSECT Constructs.
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DOI：
10.1002/cpns.30
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发表时间：
2017-07-05
影响因子：
--
通讯作者：
Deisseroth, Karl
中科院分区：
文献类型：
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作者：
Fenno, Lief E;Mattis, Joanna;Deisseroth, Karl
随着神经科学中基因编码的干预和观察工具的力量不断扩大，实验设计的界限越来越多地被在相关细胞类型中选择性表达这些工具的限制所定义。单重组酶依赖性表达系统已被广泛用作通过将重组酶依赖性病毒与表达重组酶的转基因动物组合来基于单一特征限制基因表达的手段。该方案详细说明了如何创建INTRSECT构建体并使用多种重组酶来实现所需基因靶向由多种遗传和/或拓扑特征定义的神经元子集。该方法包括病毒和转基因动物的设计和利用：这些工具具有固有的灵活性和模块化，可以以不同的组合使用，以实现所需的基因表达模式。© 2017 by John Wiley & Sons，Inc.
As the power of genetically encoded interventional and observational tools for neuroscience expands, the boundaries of experimental design are increasingly defined by limits in selectively expressing these tools in relevant cell types. Single-recombinase-dependent expression systems have been widely used as a means to restrict gene expression based on single features by combining recombinase-dependent viruses with recombinase-expressing transgenic animals. This protocol details how to create INTRSECT constructs and use multiple recombinases to achieve targeting of a desired gene to subsets of neurons that are defined by multiple genetic and/or topological features. This method includes the design and utilization of both viruses and transgenic animals: these tools are inherently flexible and modular and may be used in different combinations to achieve the desired gene expression pattern. © 2017 by John Wiley & Sons, Inc.