Viral Service: Tools for Monosynaptic Neuronal Tracing
These AAV encode tools for rabies virus-based monosynaptic tracing. These AAV can be used with modified rabies virus to perform retrograde neuronal tracing.
Note: Addgene does not distribute rabies virus.
AAV Vectors for Monosynaptic Neuronal Tracing
|52473||pAAV-synP-FLEX-splitTVA-EGFP-B19G||Can be used to complement deletion-mutant rabies virus||AAV1||Wickersham|
|100798||pAAV-syn-FLEX-splitTVA-EGFP-tTA||These AAV together can be used to complement deletion-mutant rabies virus||AAV1||Wickersham|
Introduction to Rabies Virus-based Monosynaptic Tracing
The core idea of the system is to infect the cell population of interest with a deletion-mutant tracing virus that lacks the ability to infect neighboring cells. For example, G-deleted rabies lacks the viral glycoprotein (G) gene, which is not required for the transcription or replication of the genome within infected cells, but is required for transynnaptic spread (Wickersham et al., 2007a, Wickersham et al., 2007b). Thus, G-deleted rabies can infect a single "starter" cell, but new virus cannot spread to any neighboring cells. To enable monosynaptic spread of this deletion-mutant rabies, the essential G protein is provided to the starter cell in trans, via delivery by an AAV. Since this starter cell now has all the viral genes present, the virus can spread from this cell to synaptically connected cells (meaning, it can spread to cells that are projecting to the starter cell). Once in the secondarily infected cells that lack the G protein, the virus can replicate but the newly formed viruses are unable to infect any neighboring cells. Thus, through complementation, G-deleted rabies virus can spread to monosynaptically connected cells.
To further refine the targeting of G-deleted rabies to specific starter cells, the tropism of this virus was modified so that it could only infect a genetically specified neuronal population. This is done using the envelope protein of (EnvA) of leukosis virus. EnvA can direct virus infection specifically into cells that express the TVA viral receptor. Thus, by pseudotyping the rabies virus with EnvA, only neuronal cells engineered to express TVA will be infected.
Addgene Blog: Rabies and Neuronal Tracing
Wickersham IR, Finke S, Conzelmann KK, Callaway EM. 2007a. Retrograde neuronal tracing with a deletion-mutant rabies virus. Nat Methods. 4(1):47-9. PMID: 17179932
Wickersham IR, Lyon DC, Barnard RJ, Mori T, Finke S, Conzelmann KK, Young JA, Callaway EM. 2007b. Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons. Neuron. 53(5):639-47. PMID: 17329205
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