在小鼠Th17细胞中Cis调节元件的CRISPR调节删除协议,用于体内评估效应器功能
Xiancai Zhong1, Hongmin Wu1, Guanpeng Wang1
1Department of Immunology & Theranostics, Arthur Riggs Diabetes and Metabolism Research Institute, Beckman Research Institute of the City of Hope, Duarte, CA 91010, USA.
STAR protocols
|May 21, 2025
概括
这项研究引入了CRISPR基因编辑协议,用于研究T辅助细胞17 (Th17) 中的cis调节元件. 这种方法可以在体内评估自身免疫性疾病中的基因功能,例如实验性自身免疫性脑膜炎 (EAE).
科学领域:
- 免疫学 免疫学 免疫学
- 遗传学 是一个遗传学.
- 分子生物学分子生物学
背景情况:
- 研究T辅助细胞17 (Th17) 中的 cis调节元件 (CREs) 对于了解自身免疫性疾病至关重要.
- 基因编辑的小鼠模型往往是有限的,阻碍了在实验性自身免疫脑膜炎 (EAE) 等条件下对Th17细胞功能的研究.
研究的目的:
- 提出一种CRISPR介导的CREs在小鼠Th17细胞中的删除协议.
- 为了能够在体内评估EAE模型中的Th17细胞效应器功能.
主要方法:
- 双U6gRNA结构和逆转录病毒制备.
- 病毒传递和Th17细胞分化协议.
- 在体内功能分析程序.
主要成果:
- 建立了针对Th17细胞的CRE删除的详细方案.
- 该协议在EAE模型中促进了CREs的体内功能评估.
- 这种方法克服了基因编辑小鼠稀缺所带来的局限性.
结论:
- 本次提出的基于CRISPR的协议为研究Th17细胞中的CRE功能提供了有价值的工具.
- 这种方法有助于剖析自身免疫性疾病的发病因子.
- 该协议支持在相关疾病模型中对CREs的体内功能验证.
相关概念视频
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