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2024 03 v.40 519-527
利用CRISPR/Cas9技术建立STAT1敲除细胞系及其功能验证
基金项目(Foundation): 国家自然科学基金面上项目(项目号:81971563),题目:基于汉滩和汉城病毒包膜糖蛋白保守区域设计的肾综合征出血热通用亚单位疫苗的研究;国家自然科学基金面上项目(项目号:82272330),题目:NLRC3-mTORC1轴介导的T细胞代谢重编程在HFRS急性肾损伤中的作用与机制研究;; 青年科学基金(项目号:82302525),题目:GSK3β通过磷酸化降解NP抑制汉滩病毒感染的作用和机制研究~~
邮箱(Email): gutianle1990@163.com;wuxingan@fmmu.edu.cn;
DOI: 10.13242/j.cnki.bingduxuebao.004512
中文作者单位:

空军军医大学基础医学院微生物与病原生物学教研室;延安大学生命科学学院;

摘要(Abstract):

信号转导和转录活化因子1(Signal transducer and activator of transcription 1,STAT1)是干扰素信号通路中的关键分子,在对抗病毒和其他病原体的免疫反应中发挥重要作用,构建STAT1敲除细胞系有助于研究其在病毒感染过程中的作用与机制。本研究利用CRISPR/Cas9(Clustered regularly interspaced short palindromic repeats-Cas9)基因编辑技术构建STAT1基因敲除细胞系。首先构建STAT1基因特异性打靶载体,后通过慢病毒转导人非小细胞肺癌细胞系(Human non-small cell lung cancer cells,A549),嘌呤霉素筛选稳定转导的细胞后进行单克隆化,最后对获取的单克隆细胞在分子水平上进行敲除及敲除回补验证,成功构建一株STAT1基因敲除(STAT1 gene knockout,STAT1-KO)的A549细胞,且该细胞系可维持正常细胞活性与增殖能力。以汉滩病毒(Hantaan virus,HTNV)感染验证,发现在该细胞系中病毒复制水平明显升高。本研究成功构建了一株STAT1-KO A549细胞,可为抗病毒药物和干扰素信号通路研究提供有力工具。

关键词(KeyWords): 信号转导和转录活化因子1;;CRISPR/Cas9;;基因敲除;;汉滩病毒
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基本信息:

DOI:10.13242/j.cnki.bingduxuebao.004512

中图分类号:R373

引用信息:

[1]张俊梅,王芳,应旗康等.利用CRISPR/Cas9技术建立STAT1敲除细胞系及其功能验证[J].病毒学报,2024,40(03):519-527.DOI:10.13242/j.cnki.bingduxuebao.004512.

基金信息:

国家自然科学基金面上项目(项目号:81971563),题目:基于汉滩和汉城病毒包膜糖蛋白保守区域设计的肾综合征出血热通用亚单位疫苗的研究;国家自然科学基金面上项目(项目号:82272330),题目:NLRC3-mTORC1轴介导的T细胞代谢重编程在HFRS急性肾损伤中的作用与机制研究;; 青年科学基金(项目号:82302525),题目:GSK3β通过磷酸化降解NP抑制汉滩病毒感染的作用和机制研究~~

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