Sindbis (TR339)/Venezuelan Equine Encephalitis (INH-9813) Chimeric Virus eGFP Reporter

Description: This virus is a chimeric alphavirus containing the non-structural proteins of Sindbis virus (SINV) strain TR339 and the structural proteins from Venezuelan equine encephalitis virus (VEEV) strain INH-9813. In addition, this virus expresses the reporter protein eGFP at very high levels with minimal effect on viral replicaiton or virulence. This product allows the study of VEEV INH-9813 in a non-Select Agent BSL-2 laboratory setting.

Strain History: TR339 is the consensus wild-type strain of SINV derived from the prototype AR339 isolate (Culex spp. mosquitoes, Egypt, 1952). The genomic sequence is available in NCBI. VEEV clade IC epizootic strain INH-9813 was isolated from human serum collected during the 1995 VEEV epidemic. The genomic sequence is available in NCBI.

Note: VEEV strain INH-9813 was found to contain a cell culture-adaptive mutation from glutamic acid to lysine at position 3 of the E2 protein (E3K) that confers a heparan sulfate binding phenotype (PubMed Reference).

Passage History:

• SINV TR339: Suckling mouse brain (8). References (1) and (2).
  
• VEEV INH-9813: Vero (1). Reference (3).

Production: Transfection of in vitro transcribed viral genomic RNA into baby hamster kidney (BHK) cells. No serial passages performed.

Biosafety Level: 2

Titer: 1.53 x 10⁷ PFU/mL on BHK cells.

Format: 1.0 mL/vial. Clarified and filtered cell culture supernatant frozen at -80°C. Delivered on dry ice.

QA/QC:

• Titer in PFU/mL by plaque assay
• Genome equivalents/mL by qPCR
• Whole-genome sequencing by NGS
• Mycoplasma test by PCR
• Sterility test for bacterial and fungal contamination
• Endotoxin test <10 EU/mL
• Documented expression of eGFP reporter protein
• Documentation: Experimental conditions for QC tests, documentation of all reagents used.

Reporter Virus Construction: Reporter protein coding sequences were inserted between the capsid and PE2 loci and contain the 2A-like protease sequence from the Thosea asigna virus (TaV site) at the 3’ end. Upon polyprotein translation in infected cells, the capsid autoprotease domain and the TaV protease cleave the reporter protein from the capsid, resulting in reporter expression independent of incorporation into the virus particle. For additional information, see reference.

Shipping: As this product is a Sindbis virus derivative, customers must submit a USDA APHIS permit VS 16-3 to Advanced Virology prior to order shipment. The permit and additional information are available at USDA APHIS Permits.

Production Batch Information links (downloadable):

Certificate of Analysis

NGS Results and Complete Genome Sequence

Batch Record: Detailed Production and QC Data

Product Use Policy and Material Transfer Agreement

Additional Formats: This product can be made as a purified or inactivated virus stock, or as purified viral genomic material. Many different reporter proteins and molecular tags can be incorporated. Custom variant design and production services are also available. Please contact us for additional information.

Additional Strains: If you require a strain of this virus not listed on our website, please contact us. We can produce a wide range of common strains to meet your needs.

Additional Viruses Available: If you are looking for a virus species or strain not listed on our website, please contact us with your request. Our team of experts will work with you to create the product that meets your research needs and budget.