Recombinant Mononegaviral Virus Vectors with Non-Coding Regions
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current recombinant Mononegavirales virus vectors do not achieve optimal expression levels of foreign genes and immunogenicity, as they lack the beneficial effects of non-coding regions in gene transcription units.
Innovation Solution
Incorporating 3′ and 5′ non-coding regions of Mononegavirales virus genes flanking the foreign gene within the recombinant vector, specifically between the gene start and stop codons, to enhance transcription and protein expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If foreign genes are inserted into Mononegavirales virus vectors without non-coding regions, then the vector structure remains simple, but the expression level of foreign genes is insufficient
Solution Approach 1:
The non-coding regions are pre-positioned flanking the foreign gene within the transcription unit, prepared in advance to facilitate high-level expression. This preliminary arrangement of regulatory elements ensures that when the virus infects host cells, the foreign gene is immediately subjected to optimal transcriptional control, resolving the contradiction by having the structural complexity ready beforehand rather than requiring complex regulatory mechanisms to develop during expression.
Solution Approach 2:
The non-coding regions act as intermediary elements between the viral transcription machinery and the foreign gene. These regions, containing promoter elements and other regulatory sequences, mediate the interaction between the viral RNA polymerase and the foreign gene, enabling efficient transcription without requiring direct integration of complex eukaryotic regulatory elements, thus resolving the contradiction by using viral-derived intermediaries.
2Reliability
If foreign genes are inserted into Mononegavirales virus vectors, then the vaccine can elicit immune response, but the immunogenicity is not optimal without non-coding regions
Solution Approach 1:
The invention changes the transcriptional parameters of the foreign gene by flanking it with non-coding regions containing specific promoter elements and regulatory sequences. This parameter change in the transcriptional environment transforms the foreign gene from a poorly expressed insert to a highly expressed antigen, thereby improving immunogenicity. The non-coding regions modify key transcriptional parameters such as initiation efficiency and mRNA stability without requiring complex post-translational modifications or additional viral components.
3Productivity
If non-coding regions are added to flank the foreign gene, then mRNA synthesis and protein expression increase, but the transcription unit becomes more complex
Solution Approach 1:
The non-coding regions serve multiple functions simultaneously: they provide promoter activity for transcription initiation, contain regulatory elements for mRNA processing and stability, and facilitate efficient coupling between transcription and translation. This multi-functionality allows a single structural addition (the non-coding regions) to accomplish multiple objectives that would otherwise require separate complex regulatory elements, thereby increasing productivity without proportionally increasing complexity.
Data Source
AI summary
The invention relates to a recombinant Mononegavirales virus (MV) vector comprising a foreign gene that is flanked by non-coding regions of a MV virus gene.