EHV UL43 Insertion Site Transgene Capacity
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Solution Overview
Problem
The existing EHV vector vaccines have limitations in the size and number of transgenes that can be inserted into the ORF1/3 (UL56) insertion site, necessitating the need for new and alternative transgene insertion sites to enhance vaccine capabilities.
Innovation Solution
A novel UL43 insertion site is introduced, allowing for the insertion and expression of transgenic sequences, with a partial deletion of approximately 870 bp within UL43, ensuring UL44 remains functional, and utilizing flanking regions for homologous recombination to direct the insertion of expression cassettes, enabling the expression of one or multiple antigens from the EHV vector.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If transgenes are inserted into the ORF1/3 (UL56) insertion site, then vaccine expression capability is achieved, but the size and number of transgenes that can be inserted is limited
Solution Approach 1:
The patent divides the viral genome into multiple independent insertion sites (ORF1/3 and UL43), each capable of accommodating transgenes. This segmentation allows the viral vector to carry multiple transgenes distributed across different sites, increasing overall transgene capacity without overloading a single insertion site.
Solution Approach 2:
The patent utilizes the UL43 region, which was previously unexplored for transgene insertion, adding a new dimensional space for transgene accommodation. By opening up UL43 as a functional insertion site, the patent expands the available genomic real estate beyond the traditional ORF1/3 site, enabling increased transgene load.
2Adaptability or versatility
If a partial deletion of UL43 is performed to create the insertion site, then transgene expression capability is enhanced, but viral genome integrity must be maintained
Solution Approach 1:
The patent applies a localized partial deletion in the UL43 region rather than a complete deletion or massive rearrangement. This localized approach creates sufficient space for transgene insertion while preserving the overall structural integrity and functional elements of the viral genome, maintaining reliability while enabling enhanced expression capability.
Solution Approach 2:
The patent employs a partial deletion of UL43 that is sufficient to accommodate transgenes but does not excessively disrupt viral genome integrity. This partial action strikes a balance between creating adequate insertion space and maintaining essential viral functions, ensuring both transgene expression capability and genome stability.
3Productivity
If multiple transgenes are expressed from the EHV vector, then vaccine capability is enhanced, but the complexity of managing multiple insertion sites increases
Solution Approach 1:
The patent establishes that both ORF1/3 and UL43 insertion sites can universally accommodate transgenes with similar efficiency. This universality allows the viral vector to flexibly distribute multiple transgenes across different sites without requiring site-specific optimization, simplifying the management of multiple insertions while maintaining high vaccine capability.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This solution allows for the efficient expression of one or multiple antigens from the EHV vector, reducing costs and enhancing vaccine capabilities by providing a new insertion site for transgenes, thereby overcoming the limitations of the existing ORF1/3 site.
Implementation Method 1
utilizing flanking regions for homologous recombination to direct the insertion of expression cassettes
Data Source
AI summary
The present invention relates to the field of (vector) vaccines, and especially to the novel EHV insertion site UL43. The present invention further concerns related expression cassettes and vectors, which are suitable to express genes of interest, especially antigen encoding sequences. The viral vectors of the present invention are useful for producing an immunogenic composition or vaccine.