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
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
      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.
function PdfOpen(url){ var win="toolbar=no,location=no,directories=no,status=yes,menubar=yes,scrollbars=yes,resizable=yes";,"",win); } function openWin(url,w,h){ var win="toolbar=no,location=no,directories=no,status=no,menubar=no,scrollbars=yes,resizable=no,width=" + w + ",height=" + h;,"",win); } &et=3595439A26DE3135BC601F641C7A4E3B2B28A7C8728DD896700C973E495C70FAEE5B501BA72C7C7325E1562EF4B1376EE4AAF43AEDDDD3BB1DBEBE7BD6065130D3CADE9B2E0FC4ECADF9FB2D35FE0931&pcid=&cid=&jid=&yid=0D1D160AB8016934&aid=&vid=&iid=CA4FD0336C81A37A&sid=2A3781E88AB1776F&eid=11B4E5CC8CDD3201&fileno=202001008&flag=1&is_more=0"> var my_pcid=""; var my_cid=""; var my_jid=""; var my_yid="0D1D160AB8016934"; var my_aid="";