Differences between immunodeficient mice generated by classical gene targeting and CRISPR/Cas9-mediated gene knockout |
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Authors: | " target="_blank">Jae Hoon Lee Jong-Hyung Park Tae-Wook Nam Sun-Min Seo Jun-Young Kim Han-Kyul Lee Jong Hyun Han Song Yi Park Yang-Kyu Choi Han-Woong Lee |
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Institution: | 1.Department of Biochemistry, College of Life Science and Biotechnology, Laboratory Animal Research Center,Yonsei University,Seoul,Republic of Korea;2.Department of Laboratory Animal Medicine, College of Veterinary Medicine,Konkuk University,Seoul,Republic of Korea |
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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. |
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