NATX Plasmids, for: DNA Vaccines, Gene Therapeutics, Viral Vectors andBiophamacueticals

Vaccines and Adjuvants
RIG-I Activating DNA Vaccine Vectors

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• Enhanced antigen expression in mammalian cells
• Compliance with FDA guidance
• Improved antibody response
• Improved antibody avidity
• Kanamycin or RNA-OUT (antibiotic free) selectable markers
• Improved DNA backbone for maximum plasmid production (>2.6g/L)
• Enhanced co-stimulatory RNAe™ activates RIG-1 pathway, Type I interferon production
• Optimized, chimeric promoter-intron (SV40-CMV-HTLV-1 R-U5-synthetic intron)
• Superior E. coli plasmid production yields (>2.6g/L)
• Optional TPA secretion tag

Improved: A. Interferon Response, B. Antibody Avidity, C. T cell responses
Methods to improve DNA vaccine-induced adaptive immunity are needed.  This may be accomplished using plasmid vector backbone-encoded innate immunity agonists. Retinoic-acid-inducible gene 1 (RIG-I) and melanoma differentiation-associated gene 5 (mda5) are critical cytoplasmic double stranded RNA (dsRNA) pattern receptors required for innate immune activation in response to viral infection. Activation of RIG-I and mda5 leads to type I interferon (IFN) and inflammatory cytokine production through IPS-1 activation. Since type I interferon enhances antigen-specific immune responses, we hypothesized that DNA vaccines coexpressing a RIG-I RNA agonist (eRNA) would generate superior immune responses to the encoded antigen. Indeed, expression of  RIG-I activating immunostimulatory RNA from the vector backbone (Fig 1) may be used to activate innate immunity and improve adaptive immune responses (Luke et al., 2011).


XXFig. 1

Left: RIG-1 activation of IFNβ promoter-luciferase reporter (ratio of luciferase to EGFP internal transfection control).  Synergistic RIG-1 activation by plasmid-borne eRNA VA RNA1 and eRNA 11a combination (eRNA41H). Right: Antibody avidity after prime (T=21 days) and boost (T=49 days) IM immunizations).

Left: RIG-1 activation of IFNβ promoter-luciferase reporter (ratio of luciferase to EGFP internal transfection control).  Synergistic RIG-1 activation by plasmid-borne eRNA VA RNA1 and eRNA 11a combination (eRNA41H). Right: Antibody avidity after prime (T=21 days) and boost (T=49 days) IM immunizations).
Coexpressed RIG-I Agonist Enhances Humoral Immune Response to Influenza DNA Vaccine, Luke, J.M., Simon, G.G., Soderholm, J., Errett, J.S., August, J.T., Gale, J., Hodgson, C.P., Williams, J.A. J. Virol. 85:1370-83, (2011).
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