Transgenic Nematode Phenotyping for Human Variant Function
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Solution Overview
Problem
Current transgenic animal models, such as C. elegans and zebrafish, are inefficient for accurately assessing the functional consequences of human gene variants due to limitations in expression and integration of heterologous genes, making it difficult to classify variants of uncertain significance (VUS) and identify therapeutic approaches.
Innovation Solution
Development of a transgenic animal platform using chimeric heterologous genes, where human exon coding sequences are paired with host animal intron sequences to replace native orthologs, optimizing expression and restoring function, allowing for high-throughput phenotyping and drug screening.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional transgenic animal models are used to assess human gene variant function, then the models can provide in vivo functional assessment, but the expression and integration of heterologous genes is inefficient and inaccurate
Solution Approach 1:
The patent segments the heterologous human gene into exon sequences and combines them with host animal intron sequences. This segmentation allows the human coding regions to be preserved while incorporating host regulatory elements that improve expression efficiency and integration in the transgenic animal system.
Solution Approach 2:
The patent uses host animal intron sequences as intermediaries between the human exon coding sequences and the host genome. These intron sequences facilitate proper splicing, expression, and integration of the heterologous human gene into the host animal system, resolving the incompatibility between human genes and host expression machinery.
2Measurement precision
If transgenic animal models are used for variant assessment, then functional consequences can be evaluated in native biological context, but it is difficult to classify variants of uncertain significance
Solution Approach 1:
The patent changes the structural parameters of the transgene construct by incorporating host intron sequences within the human exon coding regions. This parameter change optimizes the gene's expression characteristics in the host animal, enabling more reliable phenotypic assessment and clearer classification of variant significance based on observed functional outcomes.
3Stability of the object's composition
If human exon coding sequences are used directly without host intron sequences, then the native human gene structure is preserved, but expression and integration in host animal is inefficient
Solution Approach 1:
The patent applies local quality modification by inserting host animal intron sequences at specific locations within the human exon coding sequences. This local modification preserves the overall human gene structure and coding information while adding host-compatible regulatory elements that improve expression and integration efficiency in the transgenic animal system.
Data Source
AI summary
The present disclosure provides transgenic nematode systems for assessing function of heterologous genes, their variants and drug discovery. The transgenic nematodes contain a heterologous gene that is inserted via homologous recombination at the native locus replacing and removing the nematode ortholog, wherein expression of the heterologous gene rescues function of the removed nematode ortholog and a transgenic control animal is provided. The heterologous gene may be further modified to provide a variant, such as a human clinical variant, whereby a transgenic test animal is provided. Those transgenic test animals are used in methods to assess function of the heterologous variant and drug screens to find therapeutic candidates reversing deviant activity back to wildtype.