Two base pair deletion in IL2 receptor γ gene in NOD/SCID mice induces a highly severe immunodeficiency
Abstract Genome editing has recently emerged as a powerful tool for generating mutant mice. Small deletions of nucleotides in the target genes are frequently found in CRISPR/Cas9 mediated mutant mice. However, there are very few reports analyzing the phenotypes in small deleted mutant mice generated...
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| Format: | Article |
| Language: | English |
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BMC
2020-08-01
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| Series: | Laboratory Animal Research |
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| Online Access: | http://link.springer.com/article/10.1186/s42826-020-00048-y |
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| author | Inseon Bak Doo-Jin Kim Hyoung-Chin Kim Hye-Jun Shin Eunhye Yu Kyeong-Won Yoo Dae-Yeul Yu |
| author_facet | Inseon Bak Doo-Jin Kim Hyoung-Chin Kim Hye-Jun Shin Eunhye Yu Kyeong-Won Yoo Dae-Yeul Yu |
| author_sort | Inseon Bak |
| collection | DOAJ |
| description | Abstract Genome editing has recently emerged as a powerful tool for generating mutant mice. Small deletions of nucleotides in the target genes are frequently found in CRISPR/Cas9 mediated mutant mice. However, there are very few reports analyzing the phenotypes in small deleted mutant mice generated by CRISPR/Cas9. In this study, we generated a mutant by microinjecting sgRNAs targeting the IL2 receptor γ gene and Cas9 protein, into the cytoplasm of IVF-derived NOD.CB17/Prkdcscid/JKrb (NOD/SCID) mice embryos, and further investigated whether a 2 bp deletion of the IL2 receptor γ gene affects severe deficiency of immune cells as seen in NOD/LtSz-scid IL2 receptor γ−/− (NSG) mice. Our results show that the thymus weight of mutant mice is significantly less than that of NOD/SCID mice, whereas the spleen weight was marginally less. T and B cells in the mutant mice were severely deficient, and NK cells were almost absent. In addition, tumor growth was exceedingly increased in the mutant mice transplanted with HepG2, Raji and A549 cells, but not in nude and NOD/SCID mice. These results suggest that the NOD/SCID mice with deletion of 2 bp in the IL2 receptor γ gene shows same phenotype as NSG mice. Taken together, our data indicates that small deletions by genome editing is sufficient to generate null mutant mice. |
| first_indexed | 2024-12-12T02:59:04Z |
| format | Article |
| id | doaj.art-68674f6206884ab3ac1696a4de81d683 |
| institution | Directory Open Access Journal |
| issn | 2233-7660 |
| language | English |
| last_indexed | 2024-12-12T02:59:04Z |
| publishDate | 2020-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Laboratory Animal Research |
| spelling | doaj.art-68674f6206884ab3ac1696a4de81d6832022-12-22T00:40:40ZengBMCLaboratory Animal Research2233-76602020-08-0136111110.1186/s42826-020-00048-yTwo base pair deletion in IL2 receptor γ gene in NOD/SCID mice induces a highly severe immunodeficiencyInseon Bak0Doo-Jin Kim1Hyoung-Chin Kim2Hye-Jun Shin3Eunhye Yu4Kyeong-Won Yoo5Dae-Yeul Yu6Korea Research Institute of Bioscience and Biotechnology (KRIBB)Korea Research Institute of Bioscience and Biotechnology (KRIBB)Korea Research Institute of Bioscience and Biotechnology (KRIBB)Genome engineering laboratory, GHBIO Inc.Genome engineering laboratory, GHBIO Inc.Genome engineering laboratory, GHBIO Inc.Korea Research Institute of Bioscience and Biotechnology (KRIBB)Abstract Genome editing has recently emerged as a powerful tool for generating mutant mice. Small deletions of nucleotides in the target genes are frequently found in CRISPR/Cas9 mediated mutant mice. However, there are very few reports analyzing the phenotypes in small deleted mutant mice generated by CRISPR/Cas9. In this study, we generated a mutant by microinjecting sgRNAs targeting the IL2 receptor γ gene and Cas9 protein, into the cytoplasm of IVF-derived NOD.CB17/Prkdcscid/JKrb (NOD/SCID) mice embryos, and further investigated whether a 2 bp deletion of the IL2 receptor γ gene affects severe deficiency of immune cells as seen in NOD/LtSz-scid IL2 receptor γ−/− (NSG) mice. Our results show that the thymus weight of mutant mice is significantly less than that of NOD/SCID mice, whereas the spleen weight was marginally less. T and B cells in the mutant mice were severely deficient, and NK cells were almost absent. In addition, tumor growth was exceedingly increased in the mutant mice transplanted with HepG2, Raji and A549 cells, but not in nude and NOD/SCID mice. These results suggest that the NOD/SCID mice with deletion of 2 bp in the IL2 receptor γ gene shows same phenotype as NSG mice. Taken together, our data indicates that small deletions by genome editing is sufficient to generate null mutant mice.http://link.springer.com/article/10.1186/s42826-020-00048-yCRISPR/Cas9IL2 receptor γSmall deletionsImmunodeficient mouse |
| spellingShingle | Inseon Bak Doo-Jin Kim Hyoung-Chin Kim Hye-Jun Shin Eunhye Yu Kyeong-Won Yoo Dae-Yeul Yu Two base pair deletion in IL2 receptor γ gene in NOD/SCID mice induces a highly severe immunodeficiency Laboratory Animal Research CRISPR/Cas9 IL2 receptor γ Small deletions Immunodeficient mouse |
| title | Two base pair deletion in IL2 receptor γ gene in NOD/SCID mice induces a highly severe immunodeficiency |
| title_full | Two base pair deletion in IL2 receptor γ gene in NOD/SCID mice induces a highly severe immunodeficiency |
| title_fullStr | Two base pair deletion in IL2 receptor γ gene in NOD/SCID mice induces a highly severe immunodeficiency |
| title_full_unstemmed | Two base pair deletion in IL2 receptor γ gene in NOD/SCID mice induces a highly severe immunodeficiency |
| title_short | Two base pair deletion in IL2 receptor γ gene in NOD/SCID mice induces a highly severe immunodeficiency |
| title_sort | two base pair deletion in il2 receptor γ gene in nod scid mice induces a highly severe immunodeficiency |
| topic | CRISPR/Cas9 IL2 receptor γ Small deletions Immunodeficient mouse |
| url | http://link.springer.com/article/10.1186/s42826-020-00048-y |
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