Single-Stranded DNA Vectors for Stable Vaccine Expression

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Solution Overview

Problem

Current mRNA and dsDNA vaccines and gene therapies face challenges such as high cellular toxicity, immunogenicity, genomic integration risks, and the need for refrigeration and frequent re-administration due to mRNA instability, limiting their effectiveness and safety.

Innovation Solution

The use of single-stranded DNA (ssDNA) molecules for vaccines and gene therapies, designed with specific promoter sequences and enhancers, allows for stable expression of target sequences in cells, enabling long-term therapeutic effects without refrigeration requirements and reducing immunogenicity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If mRNA is used for vaccine or gene therapy, then adaptive immune response or therapeutic protein expression is achieved, but the mRNA requires refrigeration and is rapidly degraded leading to transient expression

Engineering Contradiction:

Improvestability of therapeutic nucleic acidVSAvoidduration of antigen or therapeutic polypeptide expression

Core Design Contradiction:

ReliabilityVSDuration of action of moving object

Solution Approach 1:

The invention changes the chemical structure parameter of the nucleic acid from RNA to single-stranded DNA. This parameter change confers greater stability to the therapeutic molecule while maintaining its ability to be transcribed into RNA and expressed as protein, thereby resolving the contradiction between stability and expression duration

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The ssDNA acts as a stable template that is copied into mRNA through transcription. This copying mechanism allows the stable DNA to produce the necessary mRNA for protein expression, combining the stability of DNA with the expressibility of RNA

Inventive Principle:

Principle #26Copying

2Reliability

If dsDNA is used for vaccine or gene therapy, then therapeutic protein expression is achieved, but cellular toxicity, immunogenicity, and risk of genomic integration occur

Engineering Contradiction:

Improvesafety profile of therapeutic nucleic acidVSAvoidcellular toxicity and immunogenicity

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The invention uses single-stranded DNA instead of double-stranded DNA, creating a locally different structural quality. This local change in strandedness reduces the activation of cGAS-STING pathways and decreases cellular toxicity and immunogenicity while maintaining therapeutic efficacy

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The ssDNA is designed to be transient and non-integrating, serving its purpose as a temporary template for transcription and then being degraded naturally. This disposable approach avoids the long-term safety risks of genomic integration associated with dsDNA

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

3Productivity

If high levels of mRNA are delivered to induce robust adaptive immune response, then vaccination efficacy is improved, but the need for frequent re-administration increases due to rapid degradation

Engineering Contradiction:

Improverobust adaptive immune responseVSAvoidfrequency of re-administration

Core Design Contradiction:

ProductivityVSLoss of time

Solution Approach 1:

The ssDNA serves as a preliminary, stable reservoir that continuously supplies mRNA through transcription. This preliminary action of providing a stable template eliminates the need for frequent re-administration while maintaining robust immune response production

Inventive Principle:

Principle #10Preliminary action

Data Source

PatentEP4663200A1Compositions and methods

Publication Date: 2025.12.17 CPTX GMBH

AI summary

Provided herein are single strands of DNA that allow for the expression of specific proteins or non-coding RNAs in a host cell or tissue. Also provided herein are vectors, lipid nanoparticles, DNA nanostructures, compositions, and vaccines comprising the single strand of DNA provided herein and uses thereof; and methods of making the same.