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URL: https://dx.doi.org/DOI:10.17504/protocols.io.84ahyse

Authors: Rosemary C. Challis, Sripriya Ravindra Kumar, Ken Y. Chan, Collin Challis, Keith Beadle, Min J. Jang, Hyun Min Kim, Pradeep S. Rajendran, John D. Tompkins, Kalyanam Shivkumar, Benjamin E. Deverman, Viviana Gradinaru

Group: Neurodegeneration Method Development Community

Summary: 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.

Affiliations: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA, Cardiac Arrhythmia Center and Neurocardiology Research Center of Excellence, University of California, Los Angeles, Los Angeles, CA, USA, Cardiac Arrhythmia Center and Neurocardiology Research Center of Excellence, University of California, Los Angeles, Los Angeles, CA, USA, Cardiac Arrhythmia Center and Neurocardiology Research Center of Excellence, University of California, Los Angeles, Los Angeles, CA, USA, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA

External URL: https://www.nature.com/articles/s41596-018-0097-3

Version: 1

Publication Date: 2019

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