Rabies Virus Vectors Construction for Optogenetic

Rabies virus (RV) is a enveloped negative strand RNA virus from the rhabdovirus family, which can selectively infect neurons rather than glia. RV vectors have become a powerful tool for circuit mapping because they infect axon terminals and then propagate only in the retrograde direction between synaptically connected neurons. This allows cross synaptic labeling of neuronal circuits across theoretically unlimited numbers of synapses. RV has been used as an effective tool to photogenetically decipher the input specific functional role of neurons projected to midbrain dopamine neurons.

The rabies virus (RVdG) method with glycoprotein (G) deletion is very useful because it can not only limit the transmission of RV by single synapses, but also limit the primary infection to genetically defined cell populations.

Creative BioMart can provide you with a series of services including RV vector design and construction, purification, production and quality verification, and provide RV vectors with high titer and purity for basic, preclinical and clinical research.

Applications of Rabies Virus Vectors

Recombinant G-deleted rabies virus can encode any gene of interest. Therefore, cell markers, function acquisition and loss analysis of specific genes, activation and silencing of specific neural circuits and monitoring of neural activities are all possible in connection defined neural networks.

For example, genetically encoded calcium indicators, voltage sensors, or glutamate sensors can monitor neural activity connecting defined neurons and/or gene targeted neurons.
Ligand gated or optically gated ion channels or pumps (channels rhodopsin, halorhodopsin and paleorhodopsin) can reversibly control neural activity to solve causality.
Photoactivated channel rhodopsin expressing neurons can reveal the synaptic output of infected neurons in slice preparation and in vivo.
CRE, FLP (flippase), tetracycline controlled trans activator (TTA) and reverse TTA (rtTA) derived from RV genome can control gene expression when used in combination with genetically engineered animals or helper viruses.

Service Content

Step	Service Description
Construction of rabies viral genome	Connecting foreign DNA to plasmid vector.
Recovery of G-deleted rabies virus from plasmids	Transfect the package cells with rabies virus genomic and helper plasmids. The package cell express both T7 polymerase and rabies virus glycoprotein.
Amplification of G-deleted rabies virus	Apply supernatant of recovered RV∆G to packages cells.
Concentration and titration of virus	Pseudotyping of RV∆G. Spin down supernatant by ultera-centrifuge Infect HEK 293T cells with serially diluted viruses

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