Recombinant Yeast for Negative-Strand RNA Virus RNP Production
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Solution Overview
Problem
Current methods for generating infectious ribonucleoprotein complexes (RNPs) of negative-strand RNA viruses, particularly non-segmented negative-strand RNA viruses, are limited by the need for mammalian cell lines that are unsuitable for large-scale production of clinical vaccine lots, and there is no established technology for producing these RNPs in yeast.
Innovation Solution
A recombinant yeast strain is developed for the expression of infectious RNPs or RNP-like particles, using DNA constructs and vectors that enable the expression of viral proteins and RNA, allowing for the production of infectious RNPs in yeast, which can be used as seeds to reproduce viral particles for vaccine components and for studying virus-host interactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If mammalian cell lines are used to generate infectious RNPs of negative-strand RNA viruses, then the production of infectious RNPs is achieved, but the suitability for large-scale production of clinical vaccine lots is poor
Solution Approach 1:
The patent introduces yeast as an intermediary host system to bridge the gap between mammalian cell culture and clinical vaccine production. Yeast serves as a safe, scalable eukaryotic platform that can express viral proteins and assemble infectious RNPs without the contamination risks and scalability limitations of mammalian cell lines, thereby enabling reliable large-scale clinical vaccine production
Solution Approach 2:
The patent creates a copy of the viral replication system within yeast cells by introducing viral RNA-dependent RNA polymerase and necessary viral proteins. This copied viral machinery enables yeast to produce authentic infectious RNPs that replicate the natural viral assembly process while maintaining the safety and scalability advantages of yeast cultivation
2Object-affected harmful factors
If yeast is used as expression cells for producing infectious RNPs, then safety and cost-effectiveness are improved, but the ability to produce infectious RNPs is not previously established
Solution Approach 1:
The patent changes the host organism parameter from mammalian cells to yeast, fundamentally altering the biological system's safety profile and cost characteristics. By modifying the host parameters to use yeast—a GRAS (Generally Recognized As Safe) organism with well-established industrial cultivation protocols—the system achieves improved safety and cost-effectiveness while maintaining RNP production capability
Solution Approach 2:
The patent segments the viral replication system into discrete components (viral RNA polymerase, nucleoprotein, phosphoprotein, and RNA genome) that can be independently expressed in yeast. This segmentation allows each component to be optimized and controlled separately, making the previously unestablished yeast-based system feasible through modular assembly of viral proteins and RNA
3Ease of manufacture
If mammalian cell lines are used for RNP production, then the production process is established, but the cost and scalability for clinical vaccine lots are limited
Solution Approach 1:
The patent makes the yeast expression system universal for producing infectious RNPs of negative-strand RNA viruses. By demonstrating that yeast can serve as a platform for multiple virus types through introduction of appropriate viral polymerases and proteins, the system achieves both ease of manufacture (using well-established yeast protocols) and high scalability (leveraging yeast's industrial fermentation capabilities)
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 method enables the high-efficiency production of infectious RNPs in yeast, providing a safe, cost-effective, and stable system for vaccine development and antiviral research, overcoming the limitations of mammalian cell lines and enabling the production of immunogenic compositions and screening for antiviral compounds.
Implementation Method 1
The present invention relates to a methodology for the generation by reverse genetics of infectious ribonucleoprotein complexes (RNPs)
Data Source
AI summary
The present invention relates to a methodology for the generation of infectious ribonucleoparticles (RNPs) of negative-strand RNA viruses, and in particular of non-segmented negative-strand RNA viruses in yeast, especially in budding yeast. Accordingly, the patent application relates to a recombinant yeast strain suitable for the rescue of infectious non-segmented negative-strand RNA virus particles or infectious virus-like particles. The invention also relates to the use of the recombinant yeast to prepare vaccine seed and to the use of the produced RNPs or RNPs-like to prepare vaccine formulations. It also concerns the use of the recombinant yeast for the screening of libraries of DNA.