Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons
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Wickersham-Production of glycoprotein.pdf
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Author(s)
Wickersham, Ian R.
Sullivan, Heather
Seung, H. Sebastian
Date Issued
March 2010
Journal
Nature Protocols
Publisher
Nature Publishing Group
Citation
Wickersham, Ian R, Heather A Sullivan, and H Sebastian Seung. “Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons.” Nature Protocols 5 (2010): 595-606. Web. 30 Nov. 2011. © 2010 Nature Publishing Group
Version
Final published version
Abstract
Recombinant rabies viruses rendered replication-deficient by the deletion of their envelope glycoprotein gene are useful tools for neuroscientists, permitting (1) extraordinarily high transgene expression levels within neurons, (2) retrograde infection of projection neurons through their axon terminals, (3) targeted infection of genetically specified neurons and (4) monosynaptic tracing of neuronal inputs. Here we present a detailed protocol for the production of high-titer and high-purity viral stocks, from initial generation of infectious virus from cDNA through amplification on complementing cell lines, pseudotyping if desired, purification by ultracentrifugation and titering. The procedure requires 3–4 weeks to complete.
MIT Department
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
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DOI of Published Version
https://doi.org/10.1038/nprot.2009.248
