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MONITTR 允许实时成像秀丽隐杆线虫中的转录和内源性蛋白。

MONITTR allows real-time imaging of transcription and endogenous proteins in C. elegans.

机构信息

The Fourth Affiliated Hospital, Zhejiang University School of Medicine , Yiwu, China.

Institute of Translational Medicine, Zhejiang University , Hangzhou, China.

出版信息

J Cell Biol. 2025 Jan 6;224(1). doi: 10.1083/jcb.202403198. Epub 2024 Oct 14.

Abstract

Maximizing cell survival under stress requires rapid and transient adjustments of RNA and protein synthesis. However, capturing these dynamic changes at both single-cell level and across an organism has been challenging. Here, we developed a system named MONITTR (MS2-embedded mCherry-based monitoring of transcription) for real-time simultaneous measurement of nascent transcripts and endogenous protein levels in C. elegans. Utilizing this system, we monitored the transcriptional bursting of fasting-induced genes and found that the epidermis responds to fasting by modulating the proportion of actively transcribing nuclei and transcriptional kinetics of individual alleles. Additionally, our findings revealed the essential roles of the transcription factors NHR-49 and HLH-30 in governing the transcriptional kinetics of fasting-induced genes under fasting. Furthermore, we tracked transcriptional dynamics during heat-shock response and ER unfolded protein response and observed rapid changes in the level of nascent transcripts under stress conditions. Collectively, our study provides a foundation for quantitatively investigating how animals spatiotemporally modulate transcription in various physiological and pathological conditions.

摘要

在压力下最大限度地提高细胞存活率需要 RNA 和蛋白质合成的快速和瞬时调整。然而,在单细胞水平和整个生物体上捕捉这些动态变化一直具有挑战性。在这里,我们开发了一种名为 MONITTR(基于 MS2 嵌入 mCherry 的转录监测)的系统,用于实时同时测量秀丽隐杆线虫中新生转录本和内源性蛋白质水平。利用该系统,我们监测了饥饿诱导基因的转录爆发,发现表皮通过调节活跃转录核的比例和单个等位基因的转录动力学来响应饥饿。此外,我们的研究结果揭示了转录因子 NHR-49 和 HLH-30 在控制饥饿诱导基因的转录动力学方面的重要作用。此外,我们在热休克反应和未折叠蛋白反应期间跟踪转录动力学,并观察到应激条件下新生转录本水平的快速变化。总的来说,我们的研究为定量研究动物如何在各种生理和病理条件下时空调节转录提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d6a/11473600/8f38df113515/JCB_202403198_Fig1.jpg

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