Engineered Cas Enzyme Platform for Precise Eukaryotic Gene Editing
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Solution Overview
Problem
Current CRISPR-Cas systems lack powerful and programmable effectors for efficient genome engineering, particularly in eukaryotic cells, necessitating the development of alternative tools for modifying nucleic acids and polynucleotides.
Innovation Solution
The development of an isolated Cas enzyme with specific amino acid sequences and fusion molecules, combined with engineered guide RNAs and vector systems, to target and alter gene expression in eukaryotic cells, including mammalian cells, by binding and cleaving target DNA strands.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If CRISPR-Cas systems are used for genome engineering in eukaryotic cells, then genome editing capability is improved, but the available effector options are limited and system complexity increases
Solution Approach 1:
The patent segments the CRISPR system into distinct functional modules: guide RNA molecules for target recognition and Cas enzymes for catalytic activity. This segmentation allows independent optimization of each component and facilitates modular assembly for different applications, resolving the complexity issue while maintaining versatility.
Solution Approach 2:
The patent develops universal Cas enzyme platforms that can perform multiple functions including DNA cleavage, base editing, and epigenetic modification. By creating multi-functional effectors, the system achieves broad adaptability across different genome engineering applications without requiring separate systems for each function, thus improving versatility while managing complexity.
2Productivity
If alternative programmable effectors are developed for eukaryotic cells, then efficiency of genome engineering is improved, but the available toolset remains insufficient
Solution Approach 1:
The patent optimizes key parameters of Cas enzymes including PAM sequence recognition, target sequence length, and catalytic activity levels. By systematically varying these parameters, the patent generates a family of effectors with different properties, thereby expanding the available toolset while maintaining high engineering efficiency across diverse applications.
Solution Approach 2:
The patent creates composite effector systems by fusing Cas enzymes with additional functional domains such as base editors, epigenetic modifiers, or fusion partners. These composite effectors provide enhanced functionality and expand the toolset available for eukaryotic genome engineering while maintaining efficient activity.
3Reliability
If current CRISPR-Cas systems are used, then DNA cleavage activity is achieved, but precision and control over gene expression alteration are limited
Solution Approach 1:
The patent introduces dynamic control mechanisms through regulatable Cas enzyme systems and inducible guide RNA expression. This allows precise temporal and spatial control over when and where DNA cleavage occurs, enhancing both the reliability of cleavage activity and the precision of gene expression control by enabling on-demand activation.
Solution Approach 2:
The patent employs intermediary molecules such as base editors and epigenetic modifiers that mediate between the Cas enzyme and the target DNA. These intermediaries provide precise control over the outcome by enabling specific modifications (e.g., base conversions, methylation) without requiring double-strand breaks, thereby improving both reliability and precision simultaneously.
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
Enables precise and efficient alteration of gene expression in eukaryotic cells, such as human cells, through the use of engineered CRISPR-Cas systems, enhancing genome engineering capabilities.
Implementation Method 1
the catalytically active domain capable of binding to a target DNA strand
Implementation Method 2
a catalytically active domain capable of cleaving the target DNA strand
Data Source
Figure 1A
Figure 1B
Figure 2A~2B
AI summary
The present application relates to the field of biomedicine, and specifically relates to a novel Cas enzyme, a system thereof and the use thereof.