Rabies Virus

  • Infection of neuronal cells
  • Trans-synaptic dissemination
  • transient transgene expression, not integrating genome
  • about 3-5 kb DNA uptake capacity
  • ss (-) RNA genome
  • Bullet shaped. 250 nm length, 80nm diameter
  • Security level S1 (if G-protein deleted)

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Rabies Viruses and its production

Rabies viruses (RABV) have the ability to spread retrograde across the synaptic cleft in a neural network. This allows the identification of neurons that are pre-synaptically linked to the first infected cell (starter cell). We use the non-pathogenic strain SAD-B19, which is also used for rabies vaccinations.

The RABV has a single-stranded RNA genome of negative polarity of about 12 kb. In the cDNA of the SAD-B19 strain, the glycoprotein gene was completely removed (G-deleted). Since transsynaptic transfer is only possible in the presence of the G protein, it must be provided in trans (by first injection with AAVs, LVs, or by transgenic G protein-expressing animals). Instead of the G protein, other proteins, such as e.g. eGFP can be expressed by the RABV.

The RABV achieves even greater safety by pseudotyping with the envelope proteins EnvA and EnvB of Avian Sarcoma and Leukosis Virus (ASLV). The infectivity of these viruses is restricted to cells carrying a non-human TVA or TVB receptor. Thus, for infection and subsequent trans-synaptic transfer, the receptor and the G protein must first be provided. The figure below describes the essential steps (modified by Wickersham et al., 2007).

Production is based on protocols developed in the labs of Edward Callaway (Salk Institute) (Osakada et al., 2013). Due to the deletion of the G protein, all subsequent work with the RABV is subject to genetic engineering safety level S1.

References:

  • Wickersham, I. R., D. C. Lyon, R. J. Barnard, T. Mori, S. Finke, K. K. Conzelmann, J. A. Young and E. M. Callaway (2007). "Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons." Neuron 53(5): 639-647.
  • Osakada, F. and E. M. Callaway (2013). "Design and generation of recombinant rabies virus vectors." Nat Protoc 8(8): 1583-1601.

Costs - Rabies virus production

Service

Costs

Discription

RABV from virus

300€

Production of RABV particles through infection with initial viruses. Purification by filtration. For in-vitro applications. 10x10ml volume. Appr. 10^5 particles/ml.

RABV from virus, for in-vivo applications

400€

Production of RABV particles through infection with initial viruses. Purification by Ultracentrifugation. 100µl volume. Appr. 10^8 particles/ml.

RABV from virus, pseudotyped

500€

Production of EnvA or EnvB pseudotyped RABV particles through infection with initial viruses. Purification by filtration. For in-vitro applications. 20x10ml volume. Appr. 10^5 particles/ml.

RABV from virus, pseudotyped, for in-vivo applications

600€

Production of EnvA or EnvB pseudotyped RABV particles through infection with initial viruses. Purification by Ultracentrifugation. 100µl volume. Appr. 10^8 particles/ml.

     

RABV from cDNA

700€

Production of RABV particles from cDNA transfection. Purification by filtration. For in-vitro applications. 10x10ml volume. Appr. 10^5 particles/ml.

RABV from cDNA, for in-vivo applications

800€

Production of RABV particles from cDNA transfection. Purification by Ultracentrifugation. 100µl volume. Appr. 10^8 particles/ml.

RABV from cDNA pseudotyped

900€

Production of EnvA or EnvB pseudotyped RABV particles from cDNA transfection. Purification by filtration. For in-vitro applications. 20x10ml volume. Appr. 10^5 particles/ml.

RABV from cDNA, pseudotyped, for in-vivo applications

1,000€

Production of EnvA or EnvB pseudotyped RABV particles from cDNA transfection. Purification by Ultracentrifugation. 100µl volume. Appr. 10^8 particles/ml.

     

RABV ready-made, pseudotyped, for in-vivo applications

50€

EnvA or EnvB pseudotyped RABV particles form our catalog. Purified by ultracentrifugation. 5µl volume. Appr. 10^8 particles/ml.