RIG-1 Inhibition for Enhanced Viral Protein Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing recombinant viral expression systems face challenges such as rapid degradation of recombinant proteins due to RIG-I mediated immune responses, leading to ineffective viral replication and transcription/translation, especially in muscle cells, and trigger immune reactions.
Innovation Solution
Transfecting muscle cells with a recombinant nucleic acid vector containing a muscle-tissue specific promoter coupled with a nucleic acid sequence to interfere with RIG-I expression, using methods like dsRNA, siRNA, or CRISPR-Cas9 to reduce RIG-I activity, combined with a viral vector encoding antigenic peptides.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If therapeutic viruses are used for transfection, then viral replication and transcription/translation occur, but immune response is triggered and RIG-I degrades viral RNA
Solution Approach 1:
The patent applies preliminary anti-action by co-administering RIG-I inhibitors (such as retinoids, small molecule inhibitors, or antisense oligonucleotides) before or concurrent with viral transfection. This pre-empts the harmful RIG-I mediated RNA degradation pathway, allowing the therapeutic virus to replicate and transcribe without being immediately degraded, thus resolving the contradiction between maintaining viral productivity and avoiding immune-mediated degradation
Solution Approach 2:
The patent introduces RIG-I inhibitors as intermediary substances that mediate between the therapeutic virus and the host immune system. These inhibitors (including retinoids like all-trans retinoic acid, small molecule compounds, or antisense oligonucleotides targeting RIG-I) act as protective intermediaries that block the harmful interaction between RIG-I and viral RNA, allowing viral replication to proceed while preventing triggering of the detrimental immune response
2Object-affected harmful factors
If E1 and E2b genes are deleted to decrease immunogenicity, then immune response is reduced, but vector gene expression decreases
Solution Approach 1:
The patent introduces RIG-I inhibitors as intermediary substances that mediate between the therapeutic virus and the host immune system. These inhibitors (including retinoids like all-trans retinoic acid, small molecule compounds, or antisense oligonucleotides targeting RIG-I) act as protective intermediaries that block the harmful interaction between RIG-I and viral RNA, allowing viral replication to proceed while preventing triggering of the detrimental immune response
Solution Approach 2:
The patent applies parameter changes by modifying the viral vector system to include RIG-I inhibition mechanisms. This could involve altering the viral genome to include sequences that inhibit RIG-I, or co-delivering RIG-I inhibitor molecules, thereby changing the immunological parameters of the viral system to allow sustained gene expression without triggering RIG-I mediated degradation
3Productivity
If high multiplicity of infection (MOI) is used, then transfection rate increases, but selective metabolic pressure increases and viral replication is disfavored
Solution Approach 1:
The patent applies preliminary anti-action by pre-treating cells with RIG-I inhibitors or pre-engineering the viral vector to include RIG-I inhibition mechanisms before transfection. This pre-empts the harmful metabolic stress response that would normally be triggered by high MOI transfection, allowing the virus to replicate efficiently even under the selective pressure imposed by high multiplicity of infection
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
Enhances recombinant protein production and reduces immune response, allowing for increased expression and presentation of viral antigens on the cell surface, thereby stimulating an effective immune response.
Implementation Method 1
transfecting into the muscle cell a nucleic acid vector comprising a muscle-tissue specific promoter and a nucleic acid sequence encoding a means of interfering with the expression of Retinoic Acid-Inducible Gene I
Implementation Method 2
using methods like dsRNA, siRNA, or CRISPR-Cas9 to reduce RIG-I activity
Implementation Method 3
a viral vector comprising a nucleic acid sequence that encodes at least one viral antigenic peptide
Implementation Method 4
allowing for increased expression and presentation of viral antigens on the cell surface
Data Source
AI summary
Viral transfection and expression of recombinant viral payload in a transfected cell is enhanced by co-expression of a means of interfering with the expression of Retinoic Acid-Inducible Gene I (RIG-1). In some embodiments, the co-expression is driven from a tissue specific promoter.