CRISPR-Cas9 sgRNA Editing of ATXN2 Gene for SCA2 Treatment
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current genome engineering techniques for treating Spinocerebellar Ataxia Type 2 (SCA2) and other ATXN2-related disorders lack precision and reproducibility, often causing severe unwanted effects due to random gene insertion, and there is a critical need for safe and effective treatments.
Innovation Solution
The use of S. pyogenes Cas9 specific single-molecule guide RNA (sgRNA) for targeted genome editing to delete or correct trinucleotide repeat expansions in the ATXN2 gene, either ex vivo or in vivo, by introducing DNA endonucleases to create single-strand or double-strand breaks, allowing for permanent changes to the genome and restoration of wild-type ATXN2 protein function.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If random insertion technologies are used to insert transgenes into living cells, then gene insertion can be achieved, but severe unwanted effects occur due to disruption of normal regulation of neighboring genes and lack of reproducibility
Solution Approach 1:
The patent introduces zinc finger nucleases (ZFNs), TALENs, and CRISPR/Cas systems as intermediary tools that mediate between the researcher's intent and the genome modification. These engineered nucleases act as precise intermediaries that recognize specific DNA sequences through programmable DNA-binding domains, enabling targeted genome editing without random insertion. The nucleases create double-strand breaks at predetermined locations, which are then repaired by cellular mechanisms to achieve the desired genetic modification.
2Manufacturing precision
If zinc finger nucleases (ZFNs), transcription activator like effector nucleases (TALENs), or CRISPR/Cas systems are used to enable specific DNA modification, then precision and reproducibility are improved, but device complexity increases
Solution Approach 1:
The patent employs segmentation by dividing the genome editing function into distinct modular components: (1) programmable DNA-binding domains (zinc finger proteins, TALE proteins, or guide RNA) that can be independently designed and optimized for different target sequences, and (2) the nuclease domain (FokI for ZFNs/TALENs or Cas9 for CRISPR) that performs the DNA cleavage. This modular architecture allows researchers to mix and match components to create customized editing systems for different genomic targets, simplifying the overall process despite the inherent complexity of the technology.
Solution Approach 2:
The patent demonstrates universality through the use of CRISPR/Cas systems, where a single Cas9 nuclease can be directed to any genomic location by simply changing the guide RNA sequence. This multi-functional platform allows the same core machinery to perform diverse genome editing tasks across different genes and organisms, reducing the need for developing entirely new systems for each application and thereby managing complexity.
3Reliability
If genome editing is performed to correct ATXN2 gene mutations, then treatment efficacy for SCA2 is improved, but off-target effects may occur
Solution Approach 1:
The patent incorporates feedback mechanisms through rigorous validation and optimization processes. Multiple sgRNAs are designed and tested to identify those with highest specificity and efficiency. Off-target effects are predicted using bioinformatic tools that analyze potential unintended binding sites throughout the genome. Experimental validation including sequencing and functional assays provide feedback on editing outcomes, allowing optimization of the editing conditions and selection of the most specific guides to minimize off-target effects while maintaining treatment efficacy.
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
This approach enables precise and reproducible editing of the ATXN2 gene, reducing or eliminating aberrant ATXN2 protein expression, thereby treating SCA2 and other related conditions by restoring normal gene function with minimal off-target effects.
Implementation Method 1
The use of S. pyogenes Cas9 specific single-molecule guide RNA (sgRNA) for targeted genome editing to delete or correct trinucleotide repeat expansions in the ATXN2 gene, either ex vivo or in vivo, by introducing DNA endonucleases to create single-strand or double-strand breaks
Data Source
Figure 1A~1B
Figure 2A
Figure 2B
AI summary
The present disclosure provides materials and methods for treating a patient with one or more conditions or disorders associated with ATXN2 whether ex vivo or in vivo. For example, the present disclosure provides materials and methods for treating a patient with Spinocerebellar ataxia type 1 (SCA2). Also provided are materials and methods for editing a ATXN2 gene in a cell by genome editing. The present disclosure also provides materials and methods for altering the contiguous genomic sequence of a ATXN2 gene in a cell. In addition, the present disclosure provides one or more gRNAs for editing a ATXN2 gene. Also provided are therapeutics comprising at least one or more gRNAs for editing a ATXN2 gene. In addition, the present disclosure provides therapeutics for treating patients with a ATXN2 related condition or disorder.