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. 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 vitro and in vivo) and in vivo delivery of the shRNAs to the brains of animals.
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Acknowledgements
Funding for these studies was provided by NIH grant HD-05751 to DWP and NHLBI grant HL-086018 to ACR.
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Ribeiro, A.C., Ågmo, A., Musatov, S., Pfaff, D.W. (2016). Silencing Estrogen Receptor-α with siRNA in the Intact Rodent Brain. In: Eyster, K.M. (eds) Estrogen Receptors. Methods in Molecular Biology, vol 1366. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3127-9_27
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DOI: https://doi.org/10.1007/978-1-4939-3127-9_27
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3126-2
Online ISBN: 978-1-4939-3127-9
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