Rabies Viruses & Psuedotyped RV Production

In a neural network, rabies viruses can spread retrogradely across synaptic clefts. It allows the identification of neurons that are pre-synaptically linked to the first infected cell (starter cell). As part of our research, we have developed the use of the non-pathogenic strain SAD-B19, which is also used for rabies vaccination, as well as CVS-N2c, which has reduced cytotoxicity and enhanced efficacy.

RABV has a single-stranded RNA genome of about 12 kb. The Rabies G protein gene was removed from SAD19 strains and CVS-N2c strains through reverse genetic approaches and replaced with a reporter gene. RABV-ΔG loses its ability to trans-synaptic, while its replication and transcription are not affected. Thus, RABV-ΔG can be used to reverse label neuronal fine morphology. Instead of the G protein, other proteins, such as e.g. eGFP can be expressed by the RABV.

A further level of safety is achieved by pseudotyping the RABV with the envelope proteins EnvA and EnvB of the Avian Sarcoma and Leukosis Virus (ASLV). These viruses are only capable of infecting cells that contain non-human TVA or TVB receptors. As long as TVA and G proteins are present, RV-ΔG can retrogradely infect upper-level neurons, enabling the labeling of trans-synaptic neural circuits.

Applications

Tracking monosynaptic input network in a specific type of neuron.
Tracking the changes in neural networks during neural development.

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