Humanized Light Chain Mice With ADAM6-Preserved Antibody Diversity
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Solution Overview
Problem
Existing transgenic mice for generating human antibodies suffer from suboptimal clonal selection processes and lack a diverse antibody repertoire due to damaged endogenous immunoglobulin loci, making them impractical for producing human therapeutics.
Innovation Solution
Genetically modified mice with reduced endogenous ADAM6 activity and ectopic ADAM6 sequences that express human immunoglobulin λ light chain variable regions, allowing rearrangement and restoration of fertility, enabling the production of diverse human antibody repertoires.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If endogenous mouse immunoglobulin loci are disabled to express human antibody sequences, then human antibody production is achieved, but fertility is lost due to disrupted ADAM6 function
Solution Approach 1:
The immunoglobulin heavy chain locus is segmented into separate functional modules: human V-D-J variable regions for antibody diversity and the mouse ADAM6 gene for fertility function. This segmentation allows independent optimization of each function without mutual interference.
Solution Approach 2:
The ADAM6 gene acts as an intermediary element within the immunoglobulin heavy chain locus, mediating between the requirement for human antibody sequences and the need for mouse fertility function. By positioning ADAM6 within this locus, the patent ensures its expression is maintained while human antibody sequences are expressed from other parts of the modified locus.
2Quantity of substance
If fully human transgenes are randomly integrated to generate human antibodies, then human antibody sequences are produced, but clonal selection becomes suboptimal due to incompatibility between human and mouse elements
Solution Approach 1:
The modified immunoglobulin heavy chain locus serves multiple functions simultaneously: it enables expression of human V-D-J variable regions for antibody diversity while maintaining mouse ADAM6 function for fertility and proper B cell development. This multi-functionality ensures that human antibody sequences are produced within a fully functional mouse immune system context, optimizing clonal selection.
3Adaptability or versatility
If endogenous ADAM6 function is disrupted to enable human immunoglobulin expression, then human antibody repertoire is expanded, but male fertility is severely reduced
Solution Approach 1:
The ADAM6 gene is preserved and positioned within the immunoglobulin heavy chain locus before human V-D-J sequences are introduced. This preliminary action ensures that ADAM6 function is maintained from the outset, preventing fertility disruption while allowing subsequent human antibody sequence expression.
Data Source
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AI summary
Non-human animals, tissues, cells, and genetic material are provided that comprise a modification of an endogenous non-human heavy chain immunoglobulin sequence and that comprise an ADAM6 activity functional in a mouse, wherein the non-human animals express a human immunoglobulin heavy chain variable domain and a cognate human immunoglobulin λ light chain variable domain.