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

Transgenic Res. 2018 Jun;27(3):241-251. doi: 10.1007/s11248-018-0069-y. Epub 2018 Mar 28.

Abstract

Immunodeficient mice are widely used for pre-clinical studies to understand various human diseases. Here, we report the generation of four immunodeficient mouse models using CRISPR/Cas9 system without inserting any foreign gene sequences such as NeoR cassettes and their characterization. By eliminating any possible effects of adding a NeoR cassette, our mouse models may allow us to better elucidate the in vivo functions of each gene. Our FVB-Rag2-/-, B6-Rag2-/-, and BALB/c-Prkdc-/- mice showed phenotypes similar to those of the earlier immunodeficient mouse models, including a lack of mature B cells and T cells and an increase in the number of CD45+DX-5+ natural killer cells. However, B6-Il2rg-/- mice had a unique phenotype, with a lack of mature B cells, increased number of T cells, and decreased number of natural killer cells. Additionally, serum immunoglobulin levels in all four immunodeficient mouse models were significantly reduced when compared to those in wild-type mice with the exception of IgM in B6-Il2rg-/- mice. These results indicate that our immunodeficient mouse models are a robust tool for in vivo studies of the immune system and will provide new insights into the variation in phenotypic outcomes resulting from different gene-targeting methodologies.

Keywords: CRISPR/Cas9; Il2rg; Immunodeficient mouse models; Prkdc; Rag2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics*
  • Disease Models, Animal
  • Gene Knockout Techniques / methods*
  • Gene Targeting / methods
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Mice, Knockout / genetics*
  • Mice, SCID / genetics*
  • Phenotype
  • T-Lymphocytes / immunology