RT Journal Article
SR Electronic
T1 Widespread and targeted gene expression by systemic AAV vectors: Production, purification, and administration
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 246405
DO 10.1101/246405
A1 Rosemary C Challis
A1 Sripriya Ravindra Kumar
A1 Ken Y Chan
A1 Collin Challis
A1 Min J Jang
A1 Pradeep S Rajendran
A1 John D Tompkins
A1 Kalyanam Shivkumar
A1 Benjamin E Deverman
A1 Viviana Gradinaru
YR 2018
UL http://biorxiv.org/content/early/2018/01/19/246405.abstract
AB We recently developed novel AAV capsids for efficient and noninvasive gene transfer across the central and peripheral nervous systems. In this protocol, we describe how to produce and systemically administer AAV-PHP viruses to label and/or genetically manipulate cells in the mouse 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 eight days, excluding the time required to assess gene expression, and can be readily adopted by laboratories with standard molecular and cell culture capabilities. We provide guidelines for experimental design and choosing 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.