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在活细胞中进行多重复合的序列成像,使用正交的化RNA胺/染料对

Ru Zheng¹, Rigumula Wu¹, Yuanchang Liu¹

¹Department of Chemistry, University of Massachusetts, Amherst, MA 01003, USA.

Nucleic acids research

|June 26, 2024

概括

研究人员开发了一种连续的化RNA成像启用传感器 (seqFRIES),用于多重活细胞成像. 这种方法允许通过连续成像和剥离光探针来检测多个目标,克服当前的局限性.

科学领域:

细胞生物学细胞生物学
分子成像学分子成像学
生物技术是生物技术.

背景情况:

多复合光成像对于理解复杂的细胞过程至关重要.
目前的方法难以在活细胞中同时检测出超过两到三个目标.

研究的目的:

引入和验证一种新的多重成像策略,seqFRIES,用于增强活细胞点检测.
为了克服与高复杂化光成像相关的技术挑战.

主要方法:

开发了一种使用基因编码的正对角化RNA吸收体的顺序化RNA成像启用传感器 (seqFRIES).
采用了连续的成像回合和细胞膜透性染料分子的快速剥离.
鉴定并优化了四个正交的化RNA吸收剂/染料对,用于活细胞成像.

主要成果:

在使用seqFRIES的活体细菌和哺乳动物细胞中展示了高度正交和多重成像.
在大约20分钟内完成了四色半定量成像过程.
成功地在单个活细胞内同时检测了关键信号分子和mRNA目标.

结论:

seqFRIES是一种经过验证的,快速且有效的多重活细胞成像策略.
开发的直角化RNA/染料对使动态和多目标检测成为可能.
这种方法有望推进活细胞成像和细胞生物学研究.

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RNA-seq

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