RNA-Based Viral Vector Delivery with Controlled Replication
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Solution Overview
Problem
Current DNA-based vectors for polynucleotide delivery face challenges such as unpredictable and irreversible genomic insertions, potential oncogenicity, immune reactions, and autoimmune-like responses due to the integration of foreign DNA, and lack of effective regulation methods.
Innovation Solution
Development of viral vectors comprising a polynucleotide encoding nucleoprotein, phosphoprotein, matrix protein, and RNA-dependent RNA polymerase, with conditionally stable fusion proteins and a glycoprotein envelope, allowing for controlled replication and transduction by utilizing stabilizing molecules, protease fusion proteins, and degron stabilizing molecules for regulated expression and safety.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If DNA-based vectors are used for polynucleotide delivery, then delivery efficiency is improved, but mutagenic risks and immune reactions increase
Solution Approach 1:
The patent extracts the problematic DNA integration function from the delivery vector by using RNA-based vesicular stomatitis virus (VSV) vectors that do not integrate into the host genome. The VSV vector delivers polynucleotides without the mutagenic insertion risks associated with DNA-based vectors like lentivirus or adeno-associated virus, thereby eliminating harmful genomic insertions while maintaining delivery efficiency
Solution Approach 2:
The patent implements periodic action through the use of conditionally stable fusion proteins containing destabilizing elements (degrons) that are temporarily stabilized by administrable molecules. This allows the viral vector to be activated only during specific treatment windows when the stabilizing molecule is present, enabling controlled, periodic expression rather than continuous expression, thus reducing immune reactions and autoimmune-like responses
2Manufacturing precision
If viral vectors are engineered with multiple control mechanisms, then regulation precision is improved, but device complexity increases
Solution Approach 1:
The patent segments the regulation system into distinct functional modules: (1) the VSV vector backbone for delivery, (2) conditionally stable fusion proteins with separable destabilizing elements and stabilizing domains, (3) administrable stabilizing molecules for activation, and (4) protease fusion proteins with controllable cleavage sites. This modular segmentation allows precise regulation at each stage while maintaining overall system manageability
Solution Approach 2:
The patent introduces intermediaries in the form of administrable stabilizing molecules that act as mediators between the external operator and the viral vector system. These molecules bind to destabilizing elements on the fusion proteins, temporarily stabilizing them to enable controlled polynucleotide expression. This intermediary mechanism provides precise temporal and spatial regulation without requiring complex genetic circuits within the vector itself
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
The solution enables safe and controlled delivery of polynucleotides, reducing mutagenic risks and immune reactions, while providing multiple layers of control for precise regulation of viral vector activity and replication, enhancing specificity and safety in biomedical applications.
Implementation Method 1
a stabilizing molecule binding domain capable of binding a stabilizing molecule, and wherein the conditionally stable fusion protein changes from a destabilized state to a stabilized state when the stabilizing molecule binding domain binds to the stabilizing molecule
Implementation Method 2
the protease is capable of cutting the first cut site and the second cut site when the protease is not bound by a protease inhibitor
Implementation Method 3
a glycoprotein binding domain of a first bridge protein is capable of binding the glycoprotein
Data Source
AI summary
Disclosed herein include methods, compositions, and systems suitable for use in delivering a polynucleotide to a target cell of a subject in need thereof. In some embodiments, a viral vector comprises a polynucleotide encoding nucleoprotein (N), phosphoprotein (P), matrix protein (M), RNA-dependent RNA polymerase (L), and one or more transgenes. The viral vector can comprise one or more of a conditionally stable fusion protein, a protease fusion protein, a degron fusion protein, and/or a glycoprotein derived of another species than the viral vector polynucleotide to enable control of viral vector transduction and/or replication.