Genotype I PRRS Virus cDNA Clone for Vaccine Development
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Solution Overview
Problem
There is a need for new infectious cDNA clones of the European genotype I PRRS virus for better understanding, comparative tests, and development of vaccines, medications, and diagnostics for Porcine Reproductive and Respiratory Syndrome (PRRS), as existing clones are limited and primarily based on the US type, with a requirement for high-yield virus production for experimental convenience.
Innovation Solution
A genotype I PRRS virus with specific nucleic acid sequences (SEQ ID NOS: 49, 56, 57, and 58) is used to create a vaccine and nucleic acid molecule, enabling the production of attenuated live viruses for preventing or treating PRRS, and a novel PRRS virus with improved properties is developed, featuring deletions in the ORF4 protein for enhanced vaccine development and immune response.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If infectious cDNA clones of European genotype I PRRS virus are developed, then understanding and comparative testing of PRRS is improved, but the complexity of virus production and manipulation increases
Solution Approach 1:
The PRRS virus genome is divided into separate cDNA fragments that are cloned into a plasmid vector system. This segmentation allows the viral genome to be constructed piece by piece in the lab, enabling controlled production while maintaining the ability to study specific genomic regions for improved understanding of PRRS.
Solution Approach 2:
A plasmid vector serves as an intermediary carrier to hold and propagate the PRRS virus cDNA sequences in bacterial or mammalian cell systems. This intermediary system simplifies virus production by using well-established molecular biology tools rather than requiring direct manipulation of the viral genome alone.
2Productivity
If high-yield virus production is achieved, then experimental convenience is improved, but the complexity of optimizing production systems increases
Solution Approach 1:
The patent optimizes various parameters including the choice of cell line (e.g., PK-15, MARC-145), transfection methods, and culture conditions to maximize virus yield from the cDNA clone. By systematically adjusting these parameters, high-yield production is achieved while keeping the overall system manageable through standardization.
3Adaptability or versatility
If novel PRRS virus variants with specific nucleic acid sequences are created, then vaccine development is improved, but the difficulty of ensuring viral safety and efficacy increases
Solution Approach 1:
Instead of working directly with live field isolates, the patent creates accurate cDNA copies of the PRRS virus genome that can be manipulated and propagated safely in the lab. These synthetic viral genomes allow vaccine development while enabling precise verification of viral sequences to ensure safety and intended efficacy.
Data Source
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AI summary
The present invention belongs to the field of animal health and provides means to study Porcine Reproductive and Respiratory Syndrome (PRRS), a viral disease affecting swine, and for the development of vaccines, therapeutics and diagnostics for the prophylaxis, treatment and diagnosis of PRRS. In a first consideration, the invention relates to a new PRRS virus variant. and, in a second consideration, to a nucleic acid sequence which comprises the genome of an infectious genotype I (EU) PRRS virus clone. Based on this, new PRRS vaccine candidates with improved properties are provided.