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Trappc11诱导型基因敲除小鼠模型的构建及鉴定
王波波1,弓梦1,温晶1,ALUSXiaoli2,李优磊1*
0
(1. 延安大学医学院生理学教研室, 延安 716000;
2. 阿尔伯特爱因斯坦医学院糖尿病研究中心, 纽约 10461
*通信作者)
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).
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