| 摘要: |
| 目的 建立转运蛋白颗粒复合物亚基11（Trappc11）诱导型基因敲除小鼠模型。方法和结果 在Trappc11基因外显子3～5的两侧分别引入loxP位点，利用CRISPR/Cas9技术获得F0代C57BL/6J小鼠；将通过PCR扩增及测序鉴定阳性的F0代C57BL/6J小鼠与C57BL/6J野生型小鼠交配、繁殖，获得F1代Trappc11flox/+小鼠；再将Trappc11flox/+小鼠与UBC-CreERT2小鼠交配，经过2代繁殖，最终获得Trappc11诱导型全身性基因敲除小鼠模型。结论 通过CRISPR/Cas9和Cre-loxP技术成功建立了Trappc11诱导型基因敲除小鼠模型，为揭示Trappc11在多器官系统疾病中的病理生理学作用提供了重要工具。 |
| 关键词: 转运蛋白颗粒复合物亚基11 基因敲除小鼠 CRISPR/Cas9系统 Cre-loxP系统 他莫昔芬 |
| DOI：10.16781/j.CN31-2187/R.20230302 |
| 投稿时间：2023-05-30修订日期：2023-08-27 |
| 基金项目:陕西省教育厅科学研究计划项目（22JK0613），陕西省自然科学基础研究计划项目（2024JC-YBQN-0949），延安大学博士科研启动项目（YDBK2021-07），陕西省高校科协青年人才托举计划（20220217）. |
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| Establishment and identification of a Trappc11 inducible knockout mouse model |
| WANG Bobo1,GONG Meng1,WEN Jing1,ALUS Xiaoli2,LI Youlei1* |
(1. Department of Physiology, Medical School of Yan'an University, Yan'an 716000, Shaanxi, China;
2. Diabetes Research Center, Albert Einstein College of Medicine, New York 10461, USA
* Corresponding author) |
| Abstract: |
| Objective To establish an inducible knockout mouse model of trafficking protein particle complex subunit 11 (Trappc11). Methods and results LoxP sites were introduced on both sides of exon 3-5 of Trappc11, and then the CRISPR/Cas9 technique was used to establish F0 C57BL/6J mice. The positive F0 generation mice were identified by polymerase chain reaction amplification and sequencing. After that, F0 positive mice were mated with C57BL/6J wild type mice to obtain F1 Trappc11flox/+ mice. And then, Trappc11flox/+ mice were mated with UBC-CreERT2 mice, and finally Trappc11 inducible systemic knockout mouse model was obtained after 2 generations. Conclusion The Trappc11 inducible knockout mouse model is established using CRISPR/Cas9 and Cre-loxP, providing an important tool for revealing the pathophysiological role of Trappc11 in multi-organ system diseases. |
| Key words: trafficking protein particle complex subunit 11 gene knockout mice CRISPR/Cas9 system Cre-loxP system tamoxifen |
