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. 2022:2418:53-61.
doi: 10.1007/978-1-0716-1920-9_4.

Immunofluorescent Verification of Silencing Estrogen Receptor α with siRNA in the Intact Rodent Brain

Ana C Ribeiro  1   2 , Mihaela A Stavarache  3 , Sergei Musatov  3 , Donald W Pfaff  4
Affiliations

Immunofluorescent Verification of Silencing Estrogen Receptor α with siRNA in the Intact Rodent Brain

Ana C Ribeiro et al. Methods Mol Biol. 2022.

Abstract

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, 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 vitro and in vivo) and in vivo delivery of the shRNAs to the brains of animals.

Keywords: Adeno-associated virus; Behavior; Estrogen receptor alpha; Mice; RNA interference; shRNA.

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