Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons
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 termin...
| Main Authors: | , , |
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2011
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| Online Access: | http://hdl.handle.net/1721.1/67322 |
| _version_ | 1826198272006422528 |
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| author | Wickersham, Ian R. Sullivan, Heather Seung, H. Sebastian |
| author2 | Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences |
| author_facet | Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Wickersham, Ian R. Sullivan, Heather Seung, H. Sebastian |
| author_sort | Wickersham, Ian R. |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T11:02:11Z |
| format | Article |
| id | mit-1721.1/67322 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:02:11Z |
| publishDate | 2011 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/673222022-09-27T16:39:38Z Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons Wickersham, Ian R. Sullivan, Heather Seung, H. Sebastian Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Wickersham, Ian R. Wickersham, Ian R. Sullivan, Heather Seung, H. Sebastian 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. Howard Hughes Medical Institute 2011-11-30T19:11:53Z 2011-11-30T19:11:53Z 2010-03 Article http://purl.org/eprint/type/JournalArticle 1750-2799 1754-2189 http://hdl.handle.net/1721.1/67322 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 en_US http://dx.doi.org/10.1038/nprot.2009.248 Nature Protocols Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Nature Publishing Group Nature |
| spellingShingle | Wickersham, Ian R. Sullivan, Heather Seung, H. Sebastian Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons |
| title | Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons |
| title_full | Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons |
| title_fullStr | Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons |
| title_full_unstemmed | Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons |
| title_short | Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons |
| title_sort | production of glycoprotein deleted rabies viruses for monosynaptic tracing and high level gene expression in neurons |
| url | http://hdl.handle.net/1721.1/67322 |
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