UTR-Engineered CRISPR Protein mRNA for Transient Expression

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Solution Overview

Problem

The application of CRISPR-Cas systems in mammalian genomes is often hampered by poor expression of Cas proteins, limiting their effectiveness in therapeutic applications such as cancer and infectious disease treatment.

Innovation Solution

The use of specific combinations of 3′ and 5′ UTR elements derived from selected genes to enhance the expression of CRISPR-associated proteins like Cas9 or Cpf1, providing transient high-expression profiles to minimize off-target effects.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional CRISPR-Cas systems are used in mammalian genomes, then genome editing function is provided, but Cas protein expression is poor

Engineering Contradiction:
ImproveCas protein expressionVSAvoidgenome editing efficiency
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent modifies the mRNA sequence parameters by incorporating specific UTR elements (5' UTR from ATP5A1, RPL32, HSD17B4, SLC7A3, NOSIP or NDUFA4; 3' UTR from GNAS, CASP1, PSMB3, ALB or RPS9) to optimize translation efficiency and protein expression levels of CRISPR-Cas proteins in mammalian cells

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent uses UTR elements as intermediary sequences that mediate between the coding region and the cellular machinery, enhancing Cas protein expression without altering the coding sequence itself

Inventive Principle:
Principle #24Intermediary (Mediator)

2Productivity

If high Cas protein expression is achieved, then genome editing efficiency is improved, but off-target effects increase

Engineering Contradiction:
Improvegenome editing efficiencyVSAvoidoff-target effects
Core Design Contradiction:
ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent employs transient high-expression profiles through optimized UTR elements, achieving high Cas protein expression for a limited duration to complete genome editing efficiently while minimizing prolonged exposure that could cause off-target effects

Inventive Principle:
Principle #19Periodic action

Data Source

PatentUS20250304976A1Nucleic acids encoding crispr-associated proteins and uses thereof
Publication Date: 2025.10.02 CUREVAC SE
  • US20250304976A1 patent drawing
  • US20250304976A1 patent drawing
  • US20250304976A1 patent drawing

AI summary

The present invention relates to the field of biomedicine, and in particular to the field of therapeutic nucleic acids. The present invention provides artificial nucleic acids, in particular RNAs, encoding CRISPR-associated proteins. A (pharmaceutical) composition and kit-of-parts comprising the same are also provided. Furthermore, the present invention relates to the artificial nucleic acid, (pharmaceutical) composition, or kit-of-parts for use in medicine, and in particular in the treatment and/or prophylaxis of diseases amenable to treatment with CRISPR-associated proteins.