Recombinant Bocavirus Vector for Helper-Free Replication

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current methods for replicating human bocavirus 1 (HBoV1) genomes are limited by the need for wild-type virus contamination and lack of specificity, which complicates gene editing and immune response enhancement, and existing vectors are inefficient for delivering therapeutic proteins to airway epithelial cells.

Innovation Solution

A recombinant BoV vector with a mutant genome that includes a 46-nucleotide minimal replication origin and mutations in non-structural proteins, allowing for helper-free replication and packaging, enabling efficient gene editing and immune response enhancement by using a predominantly negative strand genome that packages only one strand of the genome.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If wild-type virus is used for replication, then viral replication can occur, but wild-type virus contamination occurs and reduces specificity

Engineering Contradiction:

Improveviral replication efficiencyVSAvoidspecificity of gene editing

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent extracts and utilizes only the essential replication functions from the wild-type virus by providing NS1 and NP1 proteins in trans, while the viral genome lacks functional NS1 and NP1 genes. This allows replication to occur without the presence of wild-type virus, eliminating contamination while maintaining replication efficiency.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent introduces cellular proteins or proteins from other viruses as intermediaries to compensate for the defective NS1 and NP1 functions. These intermediary proteins enable viral replication and genome processing without requiring the wild-type virus, thus maintaining productivity while improving specificity.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Ease of operation

If conventional vectors are used for delivery, then gene delivery can occur, but delivery efficiency to airway epithelial cells is low

Engineering Contradiction:

Improvegene delivery capabilityVSAvoiddelivery efficiency

Core Design Contradiction:

Ease of operationVSProductivity

Solution Approach 1:

The patent modifies key parameters of the viral vector system by using a bocavirus genome with defective NS1 and NP1 genes that are complemented in trans. This parameter change creates a helper-free replication system that efficiently delivers genes to airway epithelial cells, overcoming the limitations of conventional vectors.

Inventive Principle:

Principle #35Parameter changes

3Productivity

If predominantly negative strand genome is used, then packaging efficiency improves, but genome structure complexity increases

Engineering Contradiction:

Improvepackaging efficiencyVSAvoidgenome structure

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent employs an asymmetric genome structure with a predominantly negative strand configuration. This asymmetry optimizes packaging efficiency by allowing selective packaging of one strand, while the defective NS1 and NP1 gene arrangement further enhances this asymmetric packaging mechanism without requiring wild-type virus contamination.

Inventive Principle:

Principle #4Asymmetry

Data Source

PatentUS12173305B2Cis and trans requirements for terminal resolution of human bocavirus 1

Publication Date: 2024.12.24 UNIVERSITY OF KANSAS

AI summary

A recombinant mutant BoV genome is provided, as well as methods of using the vector, e.g., to prepare helper-free virus.