Technologies
Virongy Biosciences develops cutting-edge technologies in virology and viral vectors to accelerate scientific discovery, clinical diagnostics, and disease treatment.
-
Alphavirus-based pseudoviruses are a proprietary single-cycle reporter virus platform for rapidly screening viral entry inhibitors and neutralizing antibodies. Alpha pseudoviruses consist of a virus-like particle (VLP) that encapsulates an alphavirus-derived RNA genome for rapid report expression (luciferase or GFP) in target cells (Hetrick et al., 2022). This represents a major technological advancement in the development of pseudoviruses for BSL-3 and BSL-4 level viral pathogens, allowing common biolabs to conduct rapid and robust antiviral drug screening and antibody neutralization assays. Virongy has developed a series of alpha pseudoviruses including the following viruses:
- SARS-CoV-2 and SARS-CoV: Ha-CoV-2 and Ha-CoV represents a major technology advancement in the development of SARS-CoV-2 pseudoviruses, and serves as platforms for rapid and robust quantification of neutralizing antibodies, viral mutants, and antiviral drugs (Hetrick et al., 2022; Dabbagh et al., 2021; He et al., 2021). Different from commonly used S protein pseudotyped lenti- or vesicular stomatitis virus (VSV)-psedoviruses, Ha-CoV-2 is assembled with all 4 structural proteins (S, M, N, and E) of SARS-CoV-2, and contains a reporter genome derived from an alphavirus-based vector for rapid (6 hours) and robust expression of reporter genes. Neutralization assays conducted with Ha-CoV-2 and the infectious SARS-CoV-2 showed a direct correlation (R2 = 0.87) validating that Ha-CoV-2 can serve as a surrogate of SARS-CoV-2 for neutralizing antibody quantification. SARS-CoV-2 and SARS-CoV cause respiratory coronavirus diseases and are BSL-3-level pathogens.
- Influenza A and B: Ha-IV is derived from influenza viruses, enveloped orthomyxoviruses with segmented, negative-sense RNA genomes that include influenza A and B species. Influenza viruses are major zoonotic and veterinary pathogens responsible for seasonal epidemics and sporadic pandemics, with highly pathogenic strains such as H5N1 requiring high-containment (BSL-3) facilities. The Ha-IV particles consist of the major structural proteins of influenza viruses, including hemagglutinin (HA), neuraminidase (NA), matrix proteins (M1 and M2), and nucleoprotein (NP). The Ha-IV pseudovirions encapsulate an alphavirus-derived self-amplifying RNA genome to rapidly express a reporter gene or gene of interest, enabling fast quantification of viral entry, neutralizing antibodies, and antiviral activity in a BSL-2 setting.
- Nipah Virus: Ha-NiV is derived from Nipah virus (NiV), an enveloped paramyxovirus with negative-sense and a non-segmented RNA genome. Henipaviruses are zoonotic viruses causing encephalitis and are BSL-4-level pathogens. The Ha-NiV particles consist of the four major structural proteins of the Henipavirus, the attachment glycoprotein (G), the fusion protein (F), the matrix protein (M) and the nucleocapsid protein (N). The NiV VLP encapsulates and alphavirus genome to rapidly express a reporter gene or gene of interest.
- Ebola/Marburg: Ha-EboV are derived from the Zaire Ebolavirus and Marburgvirus, members of Filoviridae with single-stranded negative-sense RNA genome. Ebola/Marburg causes. Ebolaviruses causes Ebola hemorrhagic fever and is a BSL-4-level pathogen. The Ha-EBOVV/Ha-MARV particles consist of the viral glycoprotein (GP), nucleoprotein (NP) and the VP40 matrix protein. The EBOV/MARV VLP encapsulates and alphavirus genome to rapidly express a reporter gene or gene of interest.
- Lassa mammarenavirus: Ha-LasV is derived from Lassa mammarenavirus (LASV), an arenavirus with single-stranded, bisegmented, ambisense-sense RNA genome. Lassa virus causes Lassa hemorrhagic fever and is a BSL-4-level pathogen. The Ha-LASV particle consist of the glycoprotein (GP) and the matrix protein (Z). The LASV VLP encapsulates and alphavirus genome to rapidly express a reporter gene or gene of interest.
The HIV Rev-dependent reporter cells licensed and commercialized by Virongy represent a major advancement in the development of HIV indicator cells (Wu et al., 2007). This new reporter system differs dramatically from the common LTR-based reporter cells, which rely solely on the HIV promoter, the long terminal repeat (LTR), to drive reporter expression.
The HIV Rev-dependent reporter platform are the best HIV-indicators on the market, with the highest specificity and accuracy in quantifying HIV infection. They can be used for anti-HIV drug screening, anti-HIV antibody screening, and the screening of anti-HIV compounds targeting host restriction or dependency factors (Wu et al., 2007, Curr HIV Res. 5:394).
The Exo-HEK293T enhances the production of exosomes and microvesicles, providing customers with consistent production of quality EVs that can be customized to meet individual research needs. Combine with our high-efficiency transfection reagent, Transfectin, for simple and rapid production of exosomes and EVs.
Key Features:
- High exosome and microvesicle production rate
- Customizable to fit your research needs
- Consistent production for long-term use
ExoMaxed Expression of CD9
Endo, M., Ohashi, K., and Mizuno, K. (2007). LIM kinase and slingshot are critical for neurite extension. J Biol Chem 282, 13692-13702.
Foletta, V.C., Moussi, N., Sarmiere, P.D., Bamburg, J.R., and Bernard, O. (2004). LIM kinase 1, a key regulator of actin dynamics, is widely expressed in embryonic and adult tissues. Exp Cell Res 294, 392-405.
Guo J, Wang W, Yu D, Wu Y. Spinoculation triggers dynamic actin and cofilin activity facilitating HIV-1 infection of transformed and resting CD4 T cells. J Virol. 2011; 85(19):9824-33. PubMed PMID: 21795326.
Liu, G., Xiang, Y., Guo, C., Pei, Y., Wang, Y., and Kitazato, K. (2014). Cofilin-1 is involved in regulation of actin reorganization during influenza A virus assembly and budding. Biochem Biophys Res Commun 453, 821-825.
Manetti, F. (2012). Recent Findings Confirm LIM Domain Kinases as Emerging Target Candidates for Cancer Therapy. Curr Cancer Drug Targets 12, 543-560.
Nishimura, Y., Yoshioka, K., Bernard, O., Bereczky, B., and Itoh, K. (2006). A role of LIM kinase 1/cofilin pathway in regulating endocytic trafficking of EGF receptor in human breast cancer cells. Histochem Cell Biol 126, 627-638.
Siekevitz M, Josephs SF, Dukovich M, Peffer N, Wong-Staal F, Greene WC. Activation of the HIV-1 LTR by T cell mitogens and the transactivator protein of HTLV-I. Science. 1987; 238:1575–1578.
Sweet MJ, Hume DA. RAW264 macrophages stably transfected with an HIV-1 LTR reporter gene provide a sensitive bioassay for analysis of signalling pathways in macrophages stimulated with lipopolysaccharide, TNF-alpha or taxol. J Inflamm. 1995; 45:126 –135.
Sigal A, Kim JT, Balazs AB, Dekel E, Mayo A, Milo R, et al. Cell-to-cell spread of HIV permits ongoing replication despite antiretroviral therapy. Nature. 2011; 477(7362):95-8. PubMed PMID: 21849975.
Spear M, Guo J, Turner A, Yu D, Wang W, Meltzer B, et al. HIV-1 triggers WAVE2 phosphorylation in primary CD4 T cells and macrophages, mediating Arp2/3-dependent nuclear migration. J Biol Chem. 2014; 289(10):6949-59. PubMed PMID: 24415754; PubMed Central PMCID: PMC3945356.
Sloan RD, Kuhl BD, Donahue DA, Roland A, Bar-Magen T, Wainberg MA. Transcription of preintegrated HIV-1 cDNA modulates cell surface expression of major histocompatibility complex class I via Nef. J Virol. 2011; 85(6):2828-36. PubMed PMID: 21209113.
Shuck-Lee D, Chang H, Sloan EA, Hammarskjold ML, Rekosh D. Single-nucleotide changes in the HIV Rev-response element mediate resistance to com-pounds that inhibit Rev function. J Virol. 2011; 85(8):3940-9. PubMed PMID: 21289114.
Vorster, P.J., Guo, J., Yoder, A., Wang, W., Zheng, Y., Xu, X., Yu, D., Spear, M., and Wu, Y. (2011). LIM kinase 1 modulates cortical actin and CXCR4 cycling and is activated by HIV-1 to initiate viral infection. J Biol Chem 286, 12554-12564.
Wu Y, Beddall MH, Marsh JW. Rev-dependent indicator T cell line. Current HIV Research. 2007; 5:395-403.
Yang, N., Higuchi, O., Ohashi, K., Nagata, K., Wada, A., Kangawa, K., Nishida, E., and Mizuno, K. (1998). Cofilin phosphorylation by LIM-kinase 1 and its role in Rac-mediated actin reorganization. Nature 393, 809-812.
Yi, F., Guo, J., Dabbagh, D., Spear, M., He, S., Kehn-Hall, K., Fontenot, J., Yin, Y., Bibian, M., Park, C.M., et al. (2017). Discovery of Novel Small Molecule Inhibitors of LIM Domain Kinase for Inhibiting HIV-1. J Virol. 91:e02418-16.
Yoshioka, K., Foletta, V., Bernard, O., and Itoh, K. (2003). A role for LIM kinase in cancer invasion. Proc Natl Acad Sci U S A 100, 7247-7252.
Yu D, Wang W, Yoder A, Spear M, Wu Y. The HIV envelope but not VSV glycoprotein is capable of mediating HIV latent infection of resting CD4 T cells. PLoS Pathog. 2009; 5(10):e1000633. PubMed PMID: 19851458.
Yoder A, Yu D, Dong L, Iyer SR, Xu X, Kelly J, et al. HIV envelope-CXCR4 signaling activates cofilin to overcome cortical actin restriction in resting CD4 T cells. Cell. 2008; 134(5):782-92. PubMed PMID: 18775311.