Differences between immunodeficient mice generated by classical gene targeting and CRISPR/Cas9-mediated gene...

Area of Science:

• Immunology
• Genetics
• Pre-clinical research

Background:

• Immunodeficient mice are crucial for pre-clinical disease research.
• Existing models may have confounding factors due to inserted gene sequences like NeoR cassettes.

Purpose of the Study:

• To generate and characterize novel immunodeficient mouse models using CRISPR/Cas9 without foreign gene insertions.
• To provide refined tools for in vivo gene function studies by eliminating NeoR cassette effects.

Main Methods:

• CRISPR/Cas9 gene editing was employed to create four distinct immunodeficient mouse lines.
• Phenotypic characterization included analysis of B cells, T cells, natural killer cells, and serum immunoglobulin levels.

Main Results:

• FVB-Rag2-/- , B6-Rag2-/- , and BALB/c-Prkdc-/- mice exhibited expected immunodeficient phenotypes.
• B6-Il2rg-/- mice displayed a unique phenotype: lacking mature B cells, increased T cells, and decreased natural killer cells.
• All models showed reduced serum immunoglobulin levels compared to wild-type, except for IgM in B6-Il2rg-/- mice.

Conclusions:

• The generated immunodeficient mouse models are robust tools for in vivo immune system studies.
• These models offer new insights into phenotypic variations influenced by gene-targeting methodologies.
• Eliminating foreign DNA sequences enhances the utility of these models for precise in vivo research.