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Rabies Virus (RV)

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Principle:

The Rabies virus (RV) is used for trans-monosynaptic labeling in the retrograde direction. RV belongs to the single-stranded negative sense RNA virus of the genus Lyssavirus in the family Rhabdoviridae. After RV infects the central system, it mainly labels neurons and almost no glial cells. Infected neurons have little to no significant lesions and lysis for a certain period of time (7-12 days). The replication-defective recombinant RV based on the infectious clone of the RV vaccine strain Sad-B19 has very low toxicity and can clearly label the fine morphology of neurons. It realizes the reverse cross-single-level synaptic tracing of neural network connections through the reverse complementation strategy.

The envelope glycoprotein (glycoprotein G) of RV is an essential protein for its retrograde transsynaptic function, and its receptors are widely distributed at the axon terminals, which can retrograde along the axon into the neuronal cell body to open viral replication after infection. Rabies virus for retrograde monosynaptic labeling is genetically engineered to be replication-incompetent (RV-ΔG), meaning it lacks the glycoprotein (G) necessary for further transsynaptic spread. Instead, they are pseudotyped with the avian sarcoma and leukosis virus glycoprotein EnvA (RV-EnvA-ΔG). This modification restricts the virus's ability to infect neurons unless they express a specific receptor for EnvA known as TVA, which is normally absent in mammalian cells.

To enable the rabies virus to infect only specific neurons, the TVA receptor is introduced into the targeted "starter" neurons. This is typically done by co-injecting a second virus, such as a helper AAV, which expresses the TVA receptor under the control of a neuron-specific promoter. Once the TVA receptor is expressed in the starter neurons, the RV-EnvA-ΔG can selectively infect these neurons. This ensures that the virus only infects neurons pre-engineered to express TVA, providing a high degree of specificity in selecting the initial population of neurons for labeling. Since these "starter" neurons also express the rabies glycoprotein (G), the RV spreads retrogradely across one synapse to infect presynaptic neurons. However, because the virus is replication-incompetent and lacks its own glycoprotein, it cannot spread beyond the first synapse. This results in selective labeling of only the neurons that are directly synaptically connected to the starter cells.

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RV Product List:

Biohippo offers both SAD-B19^ΔGand CVS-N2c^ΔGstrain, among which CVS-N2c^ΔG is a new glycoprotein-deletion mutant that exhibits a reduction in neuronal toxicity and a marked enhancement in transsynaptic neuronal transfer, compared to the traditional SAD-B19^ΔGstrain (Thomas R. Reardon et al., Neroresource, 2016). RV-EnvA-ΔG is from the SAD-B19^ΔG strain and RV-N2C(G)-△G is from the CVS-N2c^ΔGstrain.

Product Name	Gene Insert(s)	Color & Function	List Price (USD)
RV-ENVA-△G-EGFP	EGFP	Green; Can achieve retrograde monosynaptic labeling when used with AAV-RVG(G) and AAV-TVA helper viruses.	Inquiry
RV-ENVA-△G-DsRed	DsRed	Red; Can achieve retrograde monosynaptic labeling when used with AAV-RVG(G) and AAV-TVA helper viruses.	Inquiry
RV-ENVA-△G-GCaMP6s-DsRed	GCaMP6s, DsRed	Red. Calcium-sensitive probe. Can achieve retrograde monosynaptic labeling when used with AAV-RVG(G) and AAV-TVA helper viruses.	Inquiry
RV-ENVA-△G-mCherry	mCherry	Red; Can achieve retrograde monosynaptic labeling when used with AAV-RVG(G) and AAV-TVA helper viruses.	Inquiry
RV-N2C(G)-△G-EGFP	EGFP	Green; Non-transsynaptic	Inquiry
RV-N2C(G)-△G-DsRed	DsRed	Red; Non-transsynaptic	Inquiry
RV-N2C(G)-△G-mCherry	mCherry	Red; Non-transsynaptic	Inquiry
Customized RV			Inquiry

Case Study:

Example 1: Reverse labeling of hippocampal GABAergic neurons that simultaneously target both mPFC and ACB regions

Experimental Animals: Cre mice with inhibitory neurons (SST, PV, VIP).
Viruses Used: RV-EnvA-ΔG-GFP, AAV-DIO-TVA-hBFP, and AAV-DIO-RG.
Experimental Methods and Results: In Cre mice with inhibitory neurons (SST, PV, VIP), RV-EnvA-ΔG-GFP and Cre-dependent helper viruses AAV-DIO-TVA-hBFP and AAV-DIO-RG were injected into the mPFC brain region to achieve retrograde monosynaptic tracing. Additionally, RV-DG-DsRed, which is absorbed at the axon terminals, was injected into the ACB brain region. The virus retrogradely labeled neurons in the hippocampus that project to the ACB region, ultimately identifying GABAergic neurons in the hippocampus that simultaneously regulate both the mPFC and ACB regions (as shown in Figure 2).

Figure 2. Reverse labeling of hippocampal GABAergic neurons that simultaneously target both mPFC and ACB regions (Sunetal., Natureneuroscience, 2019). *****************************************

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Rabies Viruses & Psuedotyped RV Production

Virus and Vectors Services

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Rabies Virus (RV)

Example 1: Reverse labeling of hippocampal GABAergic neurons that simultaneously target both mPFC and ACB regions

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