Ube3a-Deficient Rat Model for Angelman Syndrome Research
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Solution Overview
Problem
Current animal models for Angelman Syndrome, such as mouse models, have limitations in accurately replicating the human condition due to potential residual isoform splicing around disrupted exons, which does not fully mimic the human disease phenotype.
Innovation Solution
Development of a UBE3A-deficient rat model with a large deletion of the entire Ube3a gene, including all isoforms and alternative promoters, using CRISPR technology to create a more accurate animal model for Angelman Syndrome, enabling comprehensive research and pre-clinical testing of therapeutics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If mouse models are used with exon disruption, then the model can be created relatively easily, but residual isoform splicing occurs that does not fully mimic the human disease phenotype
Solution Approach 1:
The patent removes the entire Ube3a gene from the rat genome using CRISPR/Cas9 technology, extracting the problematic gene entirely rather than merely disrupting exons. This complete removal eliminates residual isoform splicing while maintaining relatively straightforward model creation through targeted gene deletion.
Solution Approach 2:
The invention segments the approach to gene disruption by using CRISPR/Cas9 to target and delete the complete Ube3a gene locus, including all exons, introns, and regulatory regions. This segmentation of the gene into removable components enables complete elimination of the gene while preserving the ability to create the model through a standardized genetic engineering process.
2Manufacturing precision
If the entire Ube3a gene is deleted using CRISPR technology, then a more accurate animal model is achieved, but the complexity of the genetic modification process increases
Solution Approach 1:
The patent uses CRISPR/Cas9 as an intermediary system to achieve complete gene deletion. The Cas9 endonuclease acts as a mediator that guides RNA to the target Ube3a gene sequence, creating double-strand breaks that lead to complete gene disruption. This intermediary mechanism simplifies the process of achieving complete gene deletion compared to traditional methods.
Solution Approach 2:
The invention replaces complex traditional gene targeting methods (such as homologous recombination requiring extensive cloning and selection) with CRISPR/Cas9-mediated genome editing. This substitution of the genetic modification mechanism reduces the overall complexity while achieving complete gene deletion, as CRISPR provides a more direct and efficient pathway to gene disruption.
Data Source
AI summary
The present invention concerns non-human animals with cells having a genome that is lacking the entire E3 ubiquitin ligase (Ube3a) gene (including all isoforms and alternative promoters). These animals are useful for modeling Angelman Syndrome. The invention also includes methods for assessing the effect of an agent, such as potential therapeutics, on an animal model by exposing the animal or cells, tissues, or organs isolated therefrom, to an agent of interest.