|
| Construction and preliminary phenotypic analysis of Lag-3- / - mice |
| Received:July 22, 2019 |
| View Full Text View/Add Comment Download reader |
| DOI：10. 3969 / j.issn.1005-4847. 2020. 01. 008 |
| KeyWord:Lag-3; CRISPR/ Cas9; gene knockout; ConA |
| Author | Institution |
| 万颖寒 |
1.上海实验动物研究中心,上海 ; 2. 模式生物技术联合实验室,上海实验动物研究中心,上海 / 同济大学生命科学与技术学院,上海 |
| 慈磊 |
2. 模式生物技术联合实验室,上海实验动物研究中心,上海 / 同济大学生命科学与技术学院,上海 ; 3. 上海南方模式生物科技股份有限公司,上海 |
| 王珏 |
1.上海实验动物研究中心,上海 ; 2. 模式生物技术联合实验室,上海实验动物研究中心,上海 / 同济大学生命科学与技术学院,上海 |
| 龚慧 |
1.上海实验动物研究中心,上海 ; 2. 模式生物技术联合实验室,上海实验动物研究中心,上海 / 同济大学生命科学与技术学院,上海 |
| 万志鹏 |
3. 上海南方模式生物科技股份有限公司,上海 ; 4. 同济大学生命科学与技术学院,上海 |
| 奚骏 |
上海南方模式生物科技股份有限公司,上海 |
| 何敏珠 |
上海南方模式生物科技股份有限公司,上海 |
| 孙瑞林 |
上海南方模式生物科技股份有限公司,上海 |
| 费俭 |
2. 模式生物技术联合实验室,上海实验动物研究中心,上海 / 同济大学生命科学与技术学院,上海 ; 3. 上海南方模式生物科技股份有限公司,上海 ; 4. 同济大学生命科学与技术学院,上海 |
| 沈如凌 |
1.上海实验动物研究中心,上海 ; 2. 模式生物技术联合实验室,上海实验动物研究中心,上海 / 同济大学生命科学与技术学院,上海 |
|
| Hits: 1457 |
| Download times: 508 |
| Abstract: |
| Objective To construct lymphocyte activation gene-3 ( Lag-3)-knockout mice and analyze the influence of Lag-3 knockout on mouse phenotypes to provide an animal model for subsequent Lag-3 in vivo function research. Methods Knockout mice were constructed via CRISPR/ Cas9 technology combined with microinjection of fertilized eggs. The Lag-3-knockout mice were screened via molecular identification and further verified via RT-PCR and flow cytometry. Lag-3 gene function in vivo was studied by establishing a ConA-induced liver injury model. Results PCR and product sequencing showed that exon 2-deleted Lag-3-knockout mice were obtained. The heart, spleen, lung and macrophages from ConA-induced homozygous mice were evaluated with only Lag-3 mRNA background expression signals detected in phagocytes and lymph nodes and only very low numbers of Lag-3-positive cells detected in mouse macrophages, spleen cells and lymph nodes. Phenotypic analysis revealed that deleting the Lag-3 gene significantly reduced the number of ConA-induced CD3+T cells in the peripheral blood, bone marrow and spleen as well as ConA-induced acute liver injury. Conclusions A Lag-3-knockout mouse model is successfully constructed. In vivo functional studies during liver injury show that Lag-3 deficiency significantly alleviated ConA-induced liver damage. |
| Close |
|
|
|