Construction and preliminary phenotypic analysis of Lag-3- / - mice

1.Shanghai Laboratory Animal Research Center,Shanghai 201203,China. 2. Joint Lab for Technology of Model Organisms,Shanghai Laboratory Animal Research Center,Shanghai 201203,China / School of Life Science and Technology,Tongji University, Shanghai 200092. 3. Shanghai Model Organisms Center, Shanghai 201318. 4. School of Life Science and Technology,Tongji University, Shanghai 200092

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    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.

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  • Received:July 22,2019
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  • Online: April 01,2020
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