Nov 21, 2019
Version 1
Systemic AAV vectors for widespread and targeted gene delivery in rodents V.1
- Rosemary C. Challis1,
- Sripriya Ravindra Kumar1,
- Ken Y. Chan1,
- Collin Challis1,
- Keith Beadle1,
- Min J. Jang1,
- Hyun Min Kim1,
- Pradeep S. Rajendran2,
- John D. Tompkins2,
- Kalyanam Shivkumar2,
- Benjamin E. Deverman1,
- Viviana Gradinaru1
- 1Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA;
- 2Cardiac Arrhythmia Center and Neurocardiology Research Center of Excellence, University of California, Los Angeles, Los Angeles, CA, USA
- Neurodegeneration Method Development CommunityTech. support email: ndcn-help@chanzuckerberg.com
External link: https://www.nature.com/articles/s41596-018-0097-3
Document Citation: 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 2019. Systemic AAV vectors for widespread and targeted gene delivery in rodents. protocols.io https://dx.doi.org/10.17504/protocols.io.84ahyse
License: This is an open access document distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: November 06, 2019
Last Modified: November 21, 2019
Document Integer ID: 29538
Abstract
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.
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