CRISPR/Cas9-mediated knockout of Rag-2 causes systemic lymphopenia with hypoplastic lymphoid organs in FVB mice.
10.5625/lar.2018.34.4.166
- Author:
Joo Il KIM
1
;
Jin Sung PARK
;
Hanna KIM
;
Soo Kyung RYU
;
Jina KWAK
;
Euna KWON
;
Jun Won YUN
;
Ki Taek NAM
;
Han Woong LEE
;
Byeong Cheol KANG
Author Information
1. Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Korea. bckang@snu.ac.kr
- Publication Type:Original Article
- Keywords:
CRISPR/Cas9;
Rag-2;
histopathology;
immunodeficiency;
splenic atrophy;
thymic atrophy
- MeSH:
Animals;
Atrophy;
B-Lymphocytes;
Bone Marrow;
Genome;
Homologous Recombination;
Humans;
Immunoglobulins;
Killer Cells, Natural;
Lymph Nodes;
Lymphocytes;
Lymphopenia*;
Megakaryocytes;
Methods;
Mice*;
Mice, Knockout;
Negotiating;
Phenotype;
Receptors, Antigen, T-Cell;
Recombination, Genetic;
Sensitivity and Specificity;
Severe Combined Immunodeficiency;
Spleen;
T-Lymphocytes;
Thymus Gland
- From:Laboratory Animal Research
2018;34(4):166-175
- CountryRepublic of Korea
- Language:English
-
Abstract:
Recombination activating gene-2 (RAG-2) plays a crucial role in the development of lymphocytes by mediating recombination of T cell receptors and immunoglobulins, and loss of RAG-2 causes severe combined immunodeficiency (SCID) in humans. RAG-2 knockout mice created using homologous recombination in ES cells have served as a valuable immunodeficient platform, but concerns have persisted on the specificity of RAG-2-related phenotypes in these animals due to the limitations associated with the genome engineering method used. To precisely investigate the function of RAG-2, we recently established a new RAG-2 knockout FVB mouse line (RAG-2(−/−)) manifesting lymphopenia by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease. In this study, we further characterized their phenotypes focusing on histopathological analysis of lymphoid organs. RAG-2(−/−) mice showed no abnormality in development compared to their WT littermates for 26 weeks. At necropsy, gross examination revealed significantly smaller spleens and thymuses in RAG-2(−/−) mice, while histopathological investigation revealed hypoplastic white pulps with intact red pulps in the spleen, severe atrophy of the thymic cortex and disappearance of follicles in lymph nodes. However, no perceivable change was observed in the bone marrow. Moreover, our analyses showed a specific reduction of lymphocytes with a complete loss of mature T cells and B cells in the lymphoid organs, while natural killer cells and splenic megakaryocytes were increased in RAG-2(−/−) mice. These findings indicate that our RAG-2(−/−) mice show systemic lymphopenia with the relevant histopathological changes in the lymphoid organs, suggesting them as an improved Rag-2-related immunodeficient model.
