Monosynaptic Rabies Tracing Tools
Type: Molecular / Cellular,
Type: Other: Viral Vector,
Keywords: Monosynaptic Rabies Virus, G-Deleted Rabies, EnvA Psuedotyped Rabies, AAV Helper Virus, Circuit Tracing, TVA, Rabies Glycoprotein
Resources for Studying Neural Circuits with G-Deleted Rabies Virus
The Salk Institute maintains a viral vector core (VVC) that is partly funded by a BRAIN Initiative U24 Resource Sharing Grant (U24NS140479) titled “Resources for Studying Neural Circuits with G-Deleted Rabies Virus”. The original proposal was submitted in response to RFA-NS-23-026: Brain Initiative: Research Resource Grants for Technology Integration and Dissemination (U24). Accordingly, the work supported by this U24 is aimed to “Ensure that a diverse array of neuroscientists can access and utilize the G-deleted rabies circuit tracing platform.” Specifically, this platform involves combinations of AAV and G-deleted rabies viral vector that are used to trace the connectivity of specific cell types across the whole brain. The Salk VVC currently offers a comprehensive suite of G-deleted rabies reagents, maintains ready-to-inject stocks of rabies variants (pseudotyped with RG or EnvA) and AAV helper vectors for rapid distribution to labs throughout the US. Having a central production facility ensures consistent reagent quality and improves the rigor and reproducibility of circuit-tracing analyses.
* Resources for monosynaptic rabies tracing AAV helper viruses in many serotypes and use configurations such as Cre or Flp dependent EnvA pseudotyped
* G-deleted rabies vectors expressing a wide range of transgenes
* Availability as small aliquots to minimize cost
* Monosynaptic Rabies Tracing
* EnvA pseudotyped, G-deleted rabies vectors, along with AAV helper viruses, can be used to label the direct monosynaptic inputs to Cre- or Flp- defined cell types across the whole brain.
* G-deleted rabies can express any gene of interest, such as fluorescent proteins, genetically expressed activity indicators, or optogenetic actuators.
* AAV helper viruses in different configurations or different serotypes can be tailored to experimental aims.
Any mammalian species.
Unlike traditional anterograde or retrograde circuit tracing tools, monosynaptic rabies tracing can be used to exclusively label the direct synaptic inputs to specific cells types at a defined location within the nervous system. It is also possible to label the inputs to single neurons. By expressing various transgenes of interest, it is possible to probe or manipulate the function of neurons defined by their connectivity.
Rabies vectors used for monosynaptic tracing typically kill cell cells at about 12-14 days after infection.
Wickersham, I.R., Lyon, D.C., Barnard, R.J., Mori, T., Finke, S., Conzelmann, K.K., Young, J.A., and Callaway, E.M. (2007). Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons. Neuron 53, 639-647. 10.1016/j.neuron.2007.01.033.
Marshel, J.H., Mori, T., Nielsen, K.J., and Callaway, E.M. (2010). Targeting single neuronal networks for gene expression and cell labeling in vivo. Neuron 67, 562-574. 10.1016/j.neuron.2010.08.001.
Wall, N.R., Wickersham, I.R., Cetin, A., De La Parra, M., and Callaway, E.M. (2010). Monosynaptic circuit tracing in vivo through Cre-dependent targeting and complementation of modified rabies virus. Proceedings of the National Academy of Sciences of the United States of America 107, 21848-21853. 10.1073/pnas.1011756107.
Osakada, F., Mori, T., Cetin, A.H., Marshel, J.H., Virgen, B., and Callaway, E.M. (2011). New rabies virus variants for monitoring and manipulating activity and gene expression in defined neural circuits. Neuron 71, 617-631. 10.1016/j.neuron.2011.07.005.
Callaway, E.M., and Luo, L. (2015). Monosynaptic Circuit Tracing with Glycoprotein-Deleted Rabies Viruses. The Journal of neuroscience : the official journal of the Society for Neuroscience 35, 8979-8985. 10.1523/JNEUROSCI.0409-15.2015.
Kim, E.J., Jacobs, M.W., Ito-Cole, T., and Callaway, E.M. (2016). Improved Monosynaptic Neural Circuit Tracing Using Engineered Rabies Virus Glycoproteins. Cell reports. 10.1016/j.celrep.2016.03.067.
Kim, E.J., Zhang, Z., Huang, L., Ito-Cole, T., Jacobs, M.W., Juavinett, A.L., Senturk, G., Hu, M., Ku, M., Ecker, J.R., and Callaway, E.M. (2020). Extraction of Distinct Neuronal Cell Types from within a Genetically Continuous Population. Neuron 107, 274-282 e276. 10.1016/j.neuron.2020.04.018.
Patino, M., Lagos, W.N., Patne, N.S., Miyazaki, P.A., Bhamidipati, S.K., Collman, F., and Callaway, E.M. (2023). Postsynaptic cell type and synaptic distance do not determine efficiency of monosynaptic rabies virus spread measured at synaptic resolution. Elife 12. 10.7554/eLife.89297.
Patino, M., Rossa, M.A., Lagos, W.N., Patne, N.S., and Callaway, E.M. (2024). Transcriptomic cell-type specificity of local cortical circuits. Neuron 112, 3851-3866 e3854. 10.1016/j.neuron.2024.09.003.
Cassidy, R.M., Macias, A.V., Lagos, W.N., Ugorji, C., and Callaway, E.M. (2025). Complementary Organization of Mouse Driver and Modulator Cortico-thalamo-cortical Circuits. The Journal of neuroscience : the official journal of the Society for Neuroscience 45. 10.1523/JNEUROSCI.1167-24.2024.
* US Patent: US-20100304364-A1 Compositions and methods for monosynaptic transport Wickersham; Ian R. et al. 2010-12-02
* US Patent: US-7785874-B2 Compositions and methods for monosynaptic transport Wickersham; Ian R. et al. 2010-08-31
Edward Callaway, Professor
The Salk Institute for Biological Studies, La Jolla, CA USA
TEAM / COLLABORATOR(S)
Callaway, Edward, Professor, The Salk Institute
Naughton, John, Viral Vector Core Director, The Salk Institute
FUNDING SOURCE(S)
U24NS140479