Recombinant ORFV Vector With Multilocus Control for Polyvalent Vaccines
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Solution Overview
Problem
Existing recombinant ORFV vectors face lengthy selection processes and lack the ability to produce polyvalent vaccines, as well as insufficient control over the expression of foreign genes.
Innovation Solution
A novel recombinant ORFV vector utilizing specific insertion loci (IL 1, 2, and 3) in the ORFV genome, combined with optimized ORFV promoters (P1 and P2) for controlled expression of foreign genes, allowing for stable integration and regulated expression of multiple genes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional ORFV vectors are used for vaccine production, then the selection process is lengthy and complex, but the ability to produce polyvalent vaccines with controlled gene expression is insufficient
Solution Approach 1:
The patent divides the ORFV genome into specific functional regions (insertion loci IL-1, IL-2, IL-3) that can independently accommodate foreign genes. This segmentation allows multiple genes to be inserted at different loci simultaneously, enabling polyvalent vaccine production while maintaining organized control over each gene's expression through locus-specific promoters.
Solution Approach 2:
The patent pre-establishes specific insertion loci (IL-1, IL-2, IL-3) with defined promoter regions (P1, P2) in the ORFV genome before vaccine construction. This preliminary preparation of genomic targets eliminates the need for lengthy selection processes, as researchers can directly insert genes into these predetermined locations with known expression characteristics.
2Adaptability or versatility
If multiple foreign genes are inserted into ORFV vectors, then polyvalent vaccine capability is achieved, but control over gene expression becomes insufficient
Solution Approach 1:
The patent assigns different promoter types (P1, P2) to different insertion loci (IL-1, IL-2, IL-3) within the ORFV genome. Each promoter-local combination provides distinct expression characteristics, allowing precise control over when and how strongly each foreign gene is expressed. This local differentiation enables independent regulation of multiple genes even when inserted into the same viral vector.
Solution Approach 2:
The ORFV vector system is designed with multiple insertion loci that can each accommodate different foreign genes, making the single vector system universally applicable for producing monovalent, bivalent, or polyvalent vaccines. The same viral backbone can express any combination of antigens by selecting which loci to populate with which genes.
3Ease of manufacture
If ORFV vectors are used for gene expression, then vaccine production is enabled, but the selection and identification of recombinant viruses is time-consuming
Solution Approach 1:
The patent incorporates visual identification systems (such as fluorescent markers or colorimetric reporters) at the insertion loci that produce detectable color or fluorescence changes in cells infected with recombinant viruses. This allows rapid visual screening and identification of successfully engineered ORFV vectors without time-consuming plaque purification or molecular characterization.
Data Source
Figure 1(A)~1(C)
Figure 2
Figure 2(C)~3(C)
AI summary
The present invention relates to a recombinant Orf virus vector, a cell containing the recombinant Orf virus vector, a composition containing the recombinant Orf virus vector according to the invention and/or containing the cell according to the invention, the use of the recombinant Orf virus vector according to the invention for the production of a foreign gene. The invention also relates to a nucleic acid molecule coding for an Orf virus vector promoter.