Multipartite Lentiviral Vector System for Large Polynucleotide Delivery
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Solution Overview
Problem
Current vector systems for delivering polynucleotides into cells have limitations on the size of polynucleotides they can carry, such as AAV vectors with a packaging limit of about 5 kb and lentiviral vectors with a limit of about 10 kB, necessitating the development of technologies for larger gene delivery, including multiple vector systems and self-inactivating HIV-1 vectors.
Innovation Solution
A vector system comprising at least two polynucleotides encoding components of a macromolecular complex, specifically a multipartite cell-surface receptor, which assembles in cells to promote growth and survival, using a single lentivirus vector or two lentivirus vectors, with the assembly controlled by a ligand like rapamycin, and including transduction enhancers like anti-CD3scFv, CD86, and CD137L for efficient transduction of T cells, NK cells, and NKT cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Length of moving object
If traditional vector systems (AAV, lentiviral) are used for polynucleotide delivery, then the delivery process is simple and well-established, but the packaging size is limited (AAV: ~5 kb, Lentiviral: ~10 kB)
Solution Approach 1:
The patent divides the delivery system into multiple vector components (first vector with first polynucleotide, second vector with second polynucleotide) that can be delivered separately and then assembled within the cell. This segmentation allows each vector to stay within size limits while collectively delivering larger total genetic material.
Solution Approach 2:
The patent employs a nested structure where polynucleotides are packaged within viral capsids, which are assembled within host cells. The vectors contain packaging signals and structural proteins that form nested protective shells around the genetic material, enabling efficient delivery of larger polynucleotides through multiple smaller vector units.
2Length of moving object
If multiple vector systems are used to deliver larger genes, then the packaging size limit is overcome, but the delivery process becomes more complex
Solution Approach 1:
The patent introduces cellular assembly mechanisms as intermediaries that automatically join the delivered polynucleotides into functional macromolecular complexes. This intermediary cellular machinery simplifies the overall process by handling the complex assembly step automatically within the host cell, reducing the need for complex ex vivo manipulation.
Solution Approach 2:
The delivered polynucleotides contain self-assembly capabilities through incorporated structural proteins and packaging signals that enable automatic assembly into functional complexes within the cell. This self-service mechanism eliminates the need for complex external assembly procedures, simplifying the delivery process while achieving large gene transfer.
3Length of moving object
If self-inactivating HIV-1 vectors are used for stable transgene expression, then larger cloning capacity is achieved, but the vector system becomes more complex
Solution Approach 1:
The patent segments the vector system into multiple independent vectors, each carrying a portion of the total genetic material. This segmentation allows the use of simpler vector backbones for each individual delivery unit while collectively achieving the cloning capacity of more complex single-vector systems.
Solution Approach 2:
The patent transitions from a single-dimension vector approach (one vector carrying all genes) to a multi-dimensional approach (multiple vectors carrying different gene portions). This dimensional expansion allows the system to overcome the packaging limits of single vectors without requiring each individual vector to be overly complex.
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 system enables efficient transduction and assembly of macromolecular complexes in cells, promoting growth and survival, and effectively delivers larger genes by overcoming size limitations of traditional vector systems, with enhanced specificity and efficiency for immune cells.
Implementation Method 1
viral vector system encoding components of a macromolecular complex... A vector system comprising at least two polynucleotides, each polynucleotide comprising a polynucleotide sequence encoding a polypeptide component of a macromolecular complex
Implementation Method 2
assembly of the macromolecular complex in a cell transduced with the at least two polynucleotides promotes growth and/or survival of a cell
Implementation Method 3
the assembly of the macromolecular complex is controlled by a ligand
Data Source
AI summary
The present disclosure relates to a vector system comprising at least two polynucleotides, each polynucleotide may comprise a polynucleotide sequence encoding a polypeptide component of a macromolecular complex. Assembly of the macromolecular complex in a cell transduced with the polynucleotides may promote growth and/or survival of a cell.