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雷恩巴:用于用单细胞分辨率组合活细胞成像的光学条形码

Ruzbeh Mosadeghi¹, Daniel Foyt², Louis Sharp³

¹Department of Medicine, University of California, San Francisco, San Francisco, CA, 94143, United States of America.

bioRxiv : the preprint server for biology

|November 24, 2025

概括

我们开发了RainBar,这是一种用于活细胞成像的光学条形码系统,可实现高通量功能基因组学. 这种方法以单细胞分辨率跟踪动态细胞表型,进步我们对生物过程的理解.

科学领域:

功能性基因组学是一种功能性基因组学.
细胞成像细胞成像
分子生物学分子生物学

背景情况:

现有的高通量选方法往往错过了动态的,时间解析的细胞表型,因为它们依赖于终点测序.
活细胞成像为研究动态过程提供了潜力,但缺乏用于聚合屏幕的可扩展复合.

研究的目的:

开发一个光学条形码系统 (RainBar) 用于组合活细胞成像.
为了使功能基因组学屏幕中的动态表型能够进行高通量,单细胞分辨率的分析.
使用这个新的平台,剖析信号动态,例如NF-κB通路动力学.

主要方法:

雨吧系统利用光谱上不同的光蛋白的lentiviral联合交付,用于多重条形编码 (多达64个扰动).
先进的技术,包括单个模板病毒生产和编码多样化,以提高条形码的准确性.
基于比例的光谱分离,反向细胞质细分,以及多层感知子分类器,用于精确的条形码识别.

主要成果:

在数组和聚合格式中证明了准确的条形码识别.
成功地应用了RainBar来研究NF-κB信号动态,揭示了特定于刺激的RelA转位动力学.
在集成的CRISPRi屏幕中确定了NF-κB信号的已知和新型调节剂,包括Ino80复杂子单元和KAT2A.

结论:

雨吧是一个多功能平台,用于多重化,基于图像的活细胞功能基因组学.
该系统有助于研究动态信号架构和细胞表型.
这项技术为高分辨率和吞吐量剖析复杂的生物过程开辟了新的途径.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?光蛋白质是一种光蛋白质.活细胞成像成像技术光学条形码的使用集体选集体选集体选一个单细胞分析.

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