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使用立体定向手术技术在啮齿动物大脑中进行表观基因组编辑工具的病毒表达。

Viral Expression of Epigenome Editing Tools in Rodent Brain Using Stereotaxic Surgery Techniques.

作者信息

Hamilton Peter J, Lim Carissa J, Nestler Eric J, Heller Elizabeth A

机构信息

The Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

Department of Pharmacology and Penn Epigenetics Institute, The University of Pennsylvania, Philadelphia, PA, USA.

出版信息

Methods Mol Biol. 2018;1767:205-214. doi: 10.1007/978-1-4939-7774-1_10.

Abstract

Delivery of molecular tools for targeted epigenome editing in rodent brain can be facilitated by the use of viral vector-mediated gene transfer coupled with stereotaxic surgery techniques. Here, we describe the surgical protocol utilized by our group, which is optimized for herpes simplex virus (HSV)-mediated delivery into mouse brain. The protocol outlined herein could also be applied for delivery of adeno-associated viruses (AAV) or lentiviruses in both mice and rats. This method allows for efficient viral transgene expression and subsequent epigenome editing in rodent brain with excellent spatiotemporal control. Nearly any brain region of interest can be targeted in rodents at every stage of postnatal life. Owing to the versatility, reproducibility, and utility of this technique, it is an important method for any laboratory interested in studying the cellular, circuit, and behavioral consequences of in vivo neuroepigenome editing.

摘要

使用病毒载体介导的基因转移与立体定向手术技术相结合,可促进向啮齿动物大脑中递送用于靶向表观基因组编辑的分子工具。在此,我们描述了我们团队所采用的手术方案,该方案针对单纯疱疹病毒(HSV)介导的向小鼠大脑递送进行了优化。本文概述的方案也可应用于在小鼠和大鼠中递送腺相关病毒(AAV)或慢病毒。这种方法能够在啮齿动物大脑中实现高效的病毒转基因表达以及随后的表观基因组编辑,并具有出色的时空控制。在出生后生活的每个阶段,几乎任何感兴趣的啮齿动物脑区都可成为靶点。由于该技术的多功能性、可重复性和实用性,对于任何有兴趣研究体内神经表观基因组编辑的细胞、回路和行为后果的实验室而言,它都是一种重要的方法。

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