Generation of knockout rabbits with X-linked severe combined immunodeficiency (X-SCID) using CRISPR/Cas9

Sci Rep. 2020 Jun 19;10(1):9957. doi: 10.1038/s41598-020-66780-6.

Abstract

Severe immunodeficient mice are widely used to examine human and animal cells behaviour in vivo. However, mice are short-lived and small in size; while large animals require specific large-scale equipment. Rabbits are also commonly employed as experimental models and are larger than mice or rats, easy to handle, and suitable for long-term observational and pre-clinical studies. Herein, we sought to develop and maintain stable strains of rabbits with X-linked severe combined immunodeficiency (X-SCID) via the CRISPR/Cas9 system targeting Il2rg. Consequently, X-SCID rabbits presented immunodeficient phenotypes including the loss of T and B cells and hypoplasia of the thymus. Further, these rabbits exhibited a higher success rate with engraftments upon allogeneic transplantation of skin tissue than did wild type controls. X-SCID rabbits could be stably maintained for a minimum of four generations. These results indicate that X-SCID rabbits are effective animals for use in a non-rodent model of severe immunodeficiency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • B-Lymphocytes / immunology
  • CRISPR-Cas Systems / genetics*
  • CRISPR-Cas Systems / immunology
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Clustered Regularly Interspaced Short Palindromic Repeats / immunology
  • Female
  • Gene Knockout Techniques / methods
  • Interleukin Receptor Common gamma Subunit / genetics
  • Interleukin Receptor Common gamma Subunit / immunology
  • Rabbits
  • Skin / immunology
  • T-Lymphocytes / immunology
  • Thymus Gland / immunology
  • X-Linked Combined Immunodeficiency Diseases / genetics*
  • X-Linked Combined Immunodeficiency Diseases / immunology

Substances

  • Interleukin Receptor Common gamma Subunit