Fluorescent protein ticker tape (FPTT): Multiplexed recording of transcriptional dynamics in living cells and in vivo.

Overview

1. School of Mathematical Sciences, Peking University, Beijing, China
2. School of Life Sciences, Westlake University, Hangzhou, China
3. Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, China
4. Institute of Basic Medical Sciences, Westlake Institute for Advanced Study, Hangzhou, China
5. Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Medicine and School of Life Sciences, Westlake University, Hangzhou, Zhejiang 310024, China
6. School of Engineering, Westlake University, Hangzhou, Zhejiang 310030, China
7. College of Life Sciences, Zhejiang University, Hangzhou, China

Abstract

Recording and real-time imaging of promoter activities are critical for deciphering signaling cross-talk, but technologies for simultaneously capturing multiple transient events in living cells are lacking. Here, we design fluorescent protein–based ticker tapes (FPTT) for multiplexed, scalable, longitudinal recording of single-cell physiological activities by integrating multispectral monomeric fluorescent proteins with self-assembling protein fibers. FPTT logged dose-dependent, reversible endogenous cFos transcriptional histories in hippocampal neurons at 3-hour resolution over 8 days. We engineered FPTT variants for human nuclear factor κB (NF-κB), Janus kinase/signal transducer and activator of transcription 3 (STAT3), mechanistic target of rapamycin (mTOR), nuclear factor of activated T cells (NFAT), and adenosine 3′,5′-monophosphate (cAMP) signaling. This expanded toolset enabled quantification of cFos and NF-κB cross-talk in neurons, tracking of STAT3/cAMP dynamics during mouse liver injury, discovery of unexpected NFAT/STAT3 cross-talk, and characterization of cell cycle–dependent oscillating mTOR dynamics. Last, we achieved simultaneous analysis of four major pathways during T cell activation. FPTT provides a versatile platform to investigate transcriptional histories and signaling interplay, with broad applications in developmental biology and disease modeling.

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Data, code, and materials availability

All data and code needed to evaluate and reproduce the results in the paper are present in the paper and/or the Supplementary Materials. Raw image files with metadata, source data, raw RNA-seq datasets, and custom VBA macro scripts are publicly accessible on Zenodo (https://doi.org/10.5281/zenodo.20259137). All plasmids used in this study can be obtained from WeKwikGene (https://wekwikgene.wllsb.edu.cn/publications/0000470) under standard material transfer agreement.

Reproduced under the paper's license (CC BY-NC), from the paper cited above.

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Version 1, 27 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 5 authors, 12 MeSH terms, 3 funders, 86 references.

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This paper

Li, Z., Liu, T., Wang, Y., Xie, M., & Piatkevich, K. D. (2026). Fluorescent protein ticker tape (FPTT): Multiplexed recording of transcriptional dynamics in living cells and in vivo. Science advances, 12(36), eaef9406. https://doi.org/10.1126/sciadv.aef9406

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