Humanized MYOC Animal Models for Glaucoma Phenotype Replication
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Solution Overview
Problem
Current animal models are inadequate for replicating glaucoma phenotypes caused by MYOC mutations, which lead to trabecular meshwork stress and elevated intraocular pressure.
Innovation Solution
Development of non-human animals with a humanized MYOC locus, incorporating a human MYOC sequence to mimic MYOC mutations associated with glaucoma, and utilizing CRISPR-Cas systems for targeted gene editing and expression enhancement.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If current animal models are used, then the model simplicity is maintained, but the accuracy in replicating glaucoma phenotypes deteriorates
Solution Approach 1:
The patent applies local quality by humanizing only the specific MYOC gene locus in the animal genome while leaving the rest of the genome unchanged. This targeted approach introduces the necessary human genetic sequence to accurately replicate glaucoma phenotypes without requiring complete humanization of the entire organism, thus balancing accuracy with model simplicity.
Solution Approach 2:
The patent uses CRISPR-Cas9 gene editing technology as an intermediary tool to achieve precise modification of the MYOC locus. This intermediary system enables accurate insertion of human MYOC sequences into the animal genome, thereby improving phenotype replication accuracy without requiring complex breeding programs or multiple genetic modifications.
2Reliability
If human MYOC sequence is introduced to improve phenotype accuracy, then the relevance to human disease is improved, but the genetic stability deteriorates due to potential immunogenicity
Solution Approach 1:
By limiting humanization to only the MYOC coding sequence and essential regulatory elements rather than the entire genome, the patent maintains high human disease relevance while minimizing the introduction of foreign genetic material that could trigger immune responses or cause genomic instability.
Solution Approach 2:
The patent carefully selects and modifies specific parameters of the MYOC locus, including promoter regions, coding sequences, and polyadenylation signals, to optimize both disease relevance and genomic stability. The humanized MYOC sequence is integrated into the endogenous locus with proper regulatory elements to ensure physiological expression levels and patterns.
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
The humanized MYOC models exhibit increased MYOC expression and elevated intraocular pressure, providing a more accurate representation of glaucoma phenotypes for research and therapeutic assessment.
Implementation Method 1
modifying an endogenous MYOC locus in a non-human animal genome by: (a) introducing a double-strand break at the endogenous MYOC locus using a CRISPR-Cas system
Implementation Method 2
introducing a double-strand break at the endogenous MYOC locus using a CRISPR-Cas system
Data Source
AI summary
Non-human animal genomes, non-human animal cells, and non-human animals comprising a humanized MYOC locus and methods of making and using such non-human animal genomes, non-human animal cells, and non-human animals are provided. Non-human animal cells or non-human animals comprising a humanized MYOC locus express a human myocilin protein or a chimeric myocilin protein, fragments of which are from human myocilin. Methods are provided for using such non-human animals comprising a humanized MYOC locus to assess in vivo efficacy of human-myocilin-targeting reagents and reagents for treating glaucoma.