IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v16y2025i1d10.1038_s41467-025-58488-w.html
   My bibliography  Save this article

dFLASH; dual FLuorescent transcription factor activity sensor for histone integrated live-cell reporting and high-content screening

Author

Listed:
  • Timothy P. Allen

    (University of Adelaide)

  • Alison E. Roennfeldt

    (University of Adelaide
    University of Adelaide)

  • Moganalaxmi Reckdharajkumar

    (University of Adelaide)

  • Adrienne E. Sullivan

    (The University of Adelaide
    The University of Adelaide)

  • Miaomiao Liu

    (Griffith University)

  • Ronald J. Quinn

    (Griffith University)

  • Darryl L. Russell

    (University of Adelaide)

  • Daniel J. Peet

    (University of Adelaide)

  • Murray L. Whitelaw

    (University of Adelaide
    National Neuroscience Institute)

  • David C. Bersten

    (University of Adelaide
    University of Adelaide)

Abstract

Live-cell transcription factor (TF) activity reporting is crucial for synthetic biology, drug discovery and functional genomics. Here we present dFLASH (dual FLuorescent transcription factor Activity Sensor for Histone-integrated live-cell reporting), a modular, genome-integrated TF sensor. dFLASH homogeneously and specifically detects endogenous Hypoxia Inducible Factor (HIF) and Progesterone Receptor (PGR) activities, as well as coactivator recruitment to synthetic TFs. The dFLASH system produces dual-color nuclear fluorescence, enabling normalized, dynamic, live-cell TF activity sensing with strong signal-to-noise ratios and robust screening performance (Z’ = 0.61–0.74). We validate dFLASH for functional genomics and drug screening, demonstrating HIF regulation via CRISPRoff and application to whole-genome CRISPR KO screening. Additionally, we apply dFLASH for drug discovery, identifying HIF pathway modulators from a 1600-compound natural product library using high-content imaging. Together, this versatile platform provides a powerful tool for studying TF activity across diverse applications.

Suggested Citation

  • Timothy P. Allen & Alison E. Roennfeldt & Moganalaxmi Reckdharajkumar & Adrienne E. Sullivan & Miaomiao Liu & Ronald J. Quinn & Darryl L. Russell & Daniel J. Peet & Murray L. Whitelaw & David C. Berst, 2025. "dFLASH; dual FLuorescent transcription factor activity sensor for histone integrated live-cell reporting and high-content screening," Nature Communications, Nature, vol. 16(1), pages 1-18, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-58488-w
    DOI: 10.1038/s41467-025-58488-w
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-025-58488-w
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-025-58488-w?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-58488-w. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    We have no bibliographic references for this item. You can help adding them by using this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.