Systemic AAV vectors for widespread and targeted gene delivery in rodents
Systemic AAV vectors for widespread and targeted gene delivery in rodents
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DOI：
10.1038/s41596-018-0097-3
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发表时间：
2019-02-01
期刊：
影响因子：
14.8
通讯作者：
Gradinaru, Viviana
中科院分区：
文献类型：
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作者：
Challis, Rosemary C.;Kumar, Sripriya Ravindra;Gradinaru, Viviana
我们最近开发了腺相关病毒（AAV）衣壳，以促进有效和非侵入性的基因转移到中枢和周围神经系统。然而，产生和系统递送新型AAV变体的详细方案以前是不可用的。在本方案中，我们描述了如何生产和静脉注射aav给成年小鼠，以特异性标记和/或基因操纵神经系统和器官（包括心脏）中的细胞。该过程包括三个独立的阶段：AAV生产、静脉给药和转基因表达评估。该方案跨越8天，不包括评估基因表达所需的时间，并且可以很容易地被具有基础分子生物学，细胞培养和动物工作经验的研究人员采用。我们为实验设计和选择适合实验目的的衣壳、载物和病毒剂量提供指导。这里概述的程序适用于各种生物医学应用，从解剖和功能定位到基因表达，沉默和编辑。
We recently developed adeno-associated virus (AAV) capsids to facilitate efficient and noninvasive gene transfer to the central and peripheral nervous systems. However, a detailed protocol for generating and systemically delivering novel AAV variants was not previously available. In this protocol, we describe how to produce and intravenously administer AAVs to adult mice to specifically label and/or genetically manipulate cells in the nervous system and organs, including the heart. The procedure comprises three separate stages: AAV production, intravenous delivery, and evaluation of transgene expression. The protocol spans 8 d, excluding the time required to assess gene expression, and can be readily adopted by researchers with basic molecular biology, cell culture, and animal work experience. We provide guidelines for experimental design and choice of the capsid, cargo, and viral dose appropriate for the experimental aims. The procedures outlined here are adaptable to diverse biomedical applications, from anatomical and functional mapping to gene expression, silencing, and editing.