Modular AAV Capsid Linking for Large Gene and Protein Delivery
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Solution Overview
Problem
Current AAV-based gene delivery systems are limited by packaging size restrictions and permanent protein expression, which hinder the delivery of large or multiple genes and proteins like Cas9, leading to off-target effects.
Innovation Solution
A modular AAV-based gene and protein delivery system using peptide tags that form stable bonds, allowing AAV capsids to link together and tether proteins, expanding carrying capacity and enabling transient expression of biologically active proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If AAV vectors are used for gene delivery, then delivery efficiency is improved, but packaging size is restricted to 4.7 KB
Solution Approach 1:
The invention divides the gene delivery function into multiple AAV vectors, each carrying a portion of the total genetic material. These segmented vectors work together through complementation to deliver the complete therapeutic payload, overcoming the 4.7 KB packaging limit while maintaining high delivery efficiency of individual vectors.
Solution Approach 2:
The invention combines multiple AAV vectors into a coordinated delivery system where vectors are co-administered to target the same cells. By merging the functions of multiple vectors that each carry complementary genetic elements, the system achieves delivery of large genes that exceed the capacity of a single vector.
2Productivity
If genes are delivered via viral vectors, then gene transfer efficiency is improved, but protein expression becomes permanent
Solution Approach 1:
The invention introduces dynamic control of protein expression by incorporating inducible promoter systems and conditional expression elements into the AAV vectors. This allows the expression regime to transition from permanent to controllable, enabling researchers to induce protein expression only when needed and to regulate its duration, thereby reducing off-target effects while maintaining high gene transfer efficiency.
3Adaptability or versatility
If multiple genes are delivered separately, then each gene can be optimized, but delivery complexity increases
Solution Approach 1:
The invention creates universal AAV vector platforms with standardized elements including common promoter regions, consistent capsid structures, and uniform regulatory sequences. This universality allows different genes to be delivered using the same optimized vector backbone, reducing delivery complexity while maintaining the ability to optimize each individual gene's expression characteristics through modular genetic element selection.
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
This system doubles the carrying capacity of AAV vectors, facilitates efficient delivery of large genes and multiple genes, and reduces off-target effects by providing transient expression of proteins like Cas9.
Implementation Method 1
a peptide tag that spontaneously forms a bond (a non-limiting example of which can be a covalent bond) that is unbreakable under physiologically relevant conditions with a small globular protein
Data Source
AI summary
In one embodiment, the invention provides an Adeno-Associated Virus (AAV) comprising an exterior surface, which surface comprises one or more peptide tags that form a bond with a binding-partner, wherein the AAV is a live virus. In another embodiment, the invention provides a conjugate comprising at least one such AAV and at least one polypeptide comprising a first domain which is the binding-partner for the tag and a second domain, which is a bioactive polypeptide. In another embodiment, the invention provides a conjugate comprising at least one such AAV (first AAV) and at least one second AAV, which second AAV comprises a second exterior surface, which second exterior surface comprises at least one binding-partner for the tag or for a third linker molecule, wherein the at least one first AAV and the at least one second AAV are bound.