Manipulating gene expression in projection-specific neuronal populations using combinatorial viral approaches.
Manipulating gene expression in projection-specific neuronal populations using combinatorial viral approaches.
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DOI：
10.1002/0471142301.ns0435s65
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发表时间：
2013-01-01
影响因子：
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通讯作者：
Zweifel, Larry S
中科院分区：
文献类型：
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作者：
Gore, Bryan B;Soden, Marta E;Zweifel, Larry S
哺乳动物的大脑包含了巨大的结构和遗传复杂性，这对其功能至关重要。大脑中基因表达谱的阐明，加上大规模连接图的发展和新兴的基于病毒载体的靶向选择性基因操作方法，现在允许详细解剖基因电路接口。该方案详细介绍了如何进行组合病毒注射，以操纵连接两个大脑区域的神经元子集中的基因表达。该方法利用将逆行转导的CAV 2-Cre病毒立体定位注射到一个脑区域中，并将局部转导的Cre依赖性AAV病毒注射到另一个脑区域中。该技术广泛适用于神经回路的遗传解剖，因为它能够基于其投射靶在神经元的异质群体内选择性表达候选基因、显性阴性、荧光报告基因或遗传工具。
The mammalian brain contains tremendous structural and genetic complexity that is vital for its function. The elucidation of gene expression profiles in the brain, coupled with the development of large-scale connectivity maps and emerging viral vector-based approaches for target-selective gene manipulation, now allow for detailed dissection of gene-circuit interfaces. This protocol details how to perform combinatorial viral injections to manipulate gene expression in subsets of neurons interconnecting two brain regions. This method utilizes stereotaxic injection of a retrograde transducing CAV2-Cre virus into one brain region, combined with injection of a locally transducing Cre-dependent AAV virus into another brain region. This technique is widely applicable to the genetic dissection of neural circuitry, as it enables selective expression of candidate genes, dominant-negatives, fluorescent reporters, or genetic tools within heterogeneous populations of neurons based upon their projection targets.