计算生物学
基因传递
生物
遗传增强
基因
载体(分子生物学)
基因靶向
生物信息学
病毒学
遗传学
重组DNA
作者
Rosemary C. Challis,Sripriya Ravindra Kumar,Ken Y. Chan,Collin Challis,Keith Beadle,Min Jee Jang,Hyun Min Kim,Pradeep S. Rajendran,John D. Tompkins,Kalyanam Shivkumar,Benjamin E. Deverman,Viviana Gradinaru
出处
期刊:Nature Protocols
[Nature Portfolio]
日期:2019-01-08
卷期号:14 (2): 379-414
被引量:321
标识
DOI:10.1038/s41596-018-0097-3
摘要
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. Having developed AAV capsids that target sites throughout the body, here the authors describe how to produce and systemically administer these AAVs to rodents to label and/or genetically manipulate cells in the nervous system and visceral organs.
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