Silencing Estrogen Receptor-α with siRNA in the Intact Rodent Brain.
Silencing Estrogen Receptor-α with siRNA in the Intact Rodent Brain.
复制标题
在完整的啮齿动物大脑中用 siRNA 沉默雌激素受体-α。
DOI：
10.1007/978-1-4939-3127-9_27
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
Pfaff,DonaldW
中科院分区：
文献类型：
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作者：
Ribeiro,AnaC;Ågmo,Anders;Musatov,Sergei;Pfaff,DonaldW
在动物大脑中以化学、空间和时间特异性方式沉默基因产物表达的能力使行为神经科学领域取得了重大突破。使用这种技术，雌激素受体α（ERα）已被明确涉及小鼠的多种行为，包括性行为，攻击性行为，运动行为和母性行为。ERα已在多种脑区被鉴定，包括内侧视前区、下丘脑腹内侧和杏仁核。在本章中，我们描述了专门设计用于沉默ERα的小发夹RNA（shRNA）的产生、用于递送shRNA的腺相关病毒（AAV）载体的构建、证实ERα沉默的方法（体外和体内）以及将shRNA递送到动物脑中的体内方法。
The ability to silence the expression of gene products in a chemically, spatially, and temporally specific manner in the brains of animals has enabled key breakthroughs in the field of behavioral neuroscience. Using this technique, estrogen receptor alpha (ERα) has been specifically implicated in a multitude of behaviors in mice, including sexual, aggressive, locomotor, and maternal behaviors. ERα has been identified in a variety of brain regions, including the medial preoptic area, ventromedial hypothalamus, and amygdala. In this chapter we describe the techniques involved in the generation of the small hairpin RNAs (shRNAs) specifically designed to silence ERα, the construction of the adeno-associated viral (AAV) vector for delivery of the shRNA, the procedures to confirm the silencing of ERα (in vitroandin vivo) andin vivodelivery of the shRNAs to the brains of animals.