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用TATA-seqq进行RNA细胞下定位的现场分析的逐步协议

Xiao Jiang¹, Chu Xu¹, Lulu Hu^1,2

¹Cancer Institute, Fudan University Shanghai Cancer Center, Department of Oncology, Shanghai Key Laboratory of Medical Epigenetics, International Laboratory of Medical Epigenetics and Metabolism, Ministry of Science and Technology, Institutes of Biomedical Sciences, Shanghai Medical College, Fudan University, Shanghai, China.

|March 2, 2026

概括

我们开发了目标转录放大和测序 (TATA-seq),用于在没有膜的有机体内精确地定位RNA. 这种方法可以在特定的细胞区内进行敏感的,高准确度的RNA分析.

科学领域:

细胞生物学细胞生物学
分子生物学分子生物学
基因组学就是基因组学.

背景情况:

没有膜的器官对细胞过程至关重要,通过相分离调节生物分子.
在这些动态中心中检测RNA分布是必不可少的,但具有挑战性.
现有的方法缺乏在这些区域进行RNA分析的效率和特异性.

研究的目的:

为在没有膜的有机体内进行in situRNA分析提供一种新,高效的协议.
为了使RNA亚细胞局部化和多样性的详细分析.
为空间转录学提供一个多功能工具.

主要方法:

开发了目标转录放大和测序 (TATA-seq) 用于现场RNA检测.
利用抗体介导向和T7促进体驱动的逆转录.
集成的IgG控制用于背景减去和T7RNA聚合酶用于放大.

主要成果:

从低输入样本 (<10,000个细胞) 中实现了高保真度RNA放大.
经过证明,优化的测序指标具有<25%的重复率和~90%的映射比率.
验证了TATA-seq对各种有机体的模块性,包括核斑点和斑点.

结论:

在没有膜的有机体中,TATA-seq为RNA分析提供细胞下空间分辨率.
该协议确保高灵敏度和产量,适应各种生物条件.
这种方法推进了空间转录组学和研究细胞区内的RNA生物学.

关键词:

没有膜的有机体形成了形.RNA空间转录组学压力颗粒是一种压力颗粒.T7线性放大 T7线性放大

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