AAV Vector Packaging via Direct Repeat Removal

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Solution Overview

Problem

The production of AAV vectors encoding RdCVF and RdCVFL for treating retinal neurodegenerative disorders is limited by incomplete packaging due to direct repeated sequences within the AAV genome.

Innovation Solution

The use of AAV vectors with first and second expression cassettes that display less than 200 contiguous identical nucleotides, optimizing the production of AAV particles with a full genome.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If AAV vectors encode both RdCVF and RdCVFL with direct repeated sequences, then therapeutic functionality is improved, but packaging completeness deteriorates

Engineering Contradiction:

Improvetherapeutic functionalityVSAvoidpackaging completeness

Core Design Contradiction:

Adaptability or versatilityVSManufacturing precision

Solution Approach 1:

The patent extracts and removes the problematic direct repeated sequences from the AAV genome while retaining the essential therapeutic genes. Specifically, the invention eliminates the direct repeats between RdCVF and RdCVFL expression cassettes that cause packaging defects, thereby resolving the contradiction between maintaining therapeutic functionality and achieving complete packaging.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent changes the genomic parameters by altering the sequence arrangement and reducing the length of direct repeated sequences. By modifying the nucleotide sequence structure (reducing repeat length from hundreds to minimal sequences), the invention enables complete packaging while preserving the therapeutic expression of both RdCVF and RdCVFL proteins.

Inventive Principle:

Principle #35Parameter changes

2Productivity

If AAV genome contains long direct repeated sequences, then gene expression is enhanced, but vector production is compromised

Engineering Contradiction:

Improvegene expressionVSAvoidvector production

Core Design Contradiction:

ProductivityVSEase of manufacture

Solution Approach 1:

The invention extracts and removes the problematic direct repeated sequences that interfere with vector production. By eliminating these sequences while maintaining the essential coding regions for RdCVF and RdCVFL, the patent achieves both effective gene expression and successful vector production without manufacturing complications.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent uses alternative sequence arrangements that replicate the therapeutic functionality without the harmful direct repeats. By creating modified expression cassettes that achieve similar gene expression outcomes through different sequence configurations, the invention enables easy vector production while maintaining productivity.

Inventive Principle:

Principle #26Copying

3Reliability

If AAV vectors are designed for GMP compliance, then therapeutic quality is improved, but packaging efficiency deteriorates

Engineering Contradiction:

Improvetherapeutic qualityVSAvoidpackaging efficiency

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The patent changes the genomic parameters by optimizing the sequence structure to eliminate direct repeats that cause packaging inefficiency. By reducing the length and number of direct repeated sequences between expression cassettes, the invention achieves complete packaging efficiency while maintaining GMP compliance and therapeutic quality standards.

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS20250059563A1Constructs comprising neuronal viability factors and uses thereof

Publication Date: 2025.02.20 SPARINGVISION

AI summary

The present invention relates to improved constructs comprising the short and long Rod-Derived Cone Viability Factors and to methods for treating retinal degenerative diseases.