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A genetically encoded probe for imaging nascent and mature HA-tagged proteins in vivo

Author

Listed:
  • Ning Zhao

    (Colorado State University)

  • Kouta Kamijo

    (Tokyo Institute of Technology)

  • Philip D. Fox

    (Colorado State University)

  • Haruka Oda

    (Tokyo Institute of Technology)

  • Tatsuya Morisaki

    (Colorado State University)

  • Yuko Sato

    (Tokyo Institute of Technology
    Tokyo Institute of Technology)

  • Hiroshi Kimura

    (Tokyo Institute of Technology
    Tokyo Institute of Technology
    Tokyo Institute of Technology)

  • Timothy J. Stasevich

    (Colorado State University
    Tokyo Institute of Technology)

Abstract

To expand the toolbox of imaging in living cells, we have engineered a single-chain variable fragment binding the linear HA epitope with high affinity and specificity in vivo. The resulting probe, called the HA frankenbody, can light up in multiple colors HA-tagged nuclear, cytoplasmic, membrane, and mitochondrial proteins in diverse cell types. The HA frankenbody also enables state-of-the-art single-molecule experiments in living cells, which we demonstrate by tracking single HA-tagged histones in U2OS cells and single mRNA translation dynamics in both U2OS cells and neurons. Together with the SunTag, we also track two mRNA species simultaneously to demonstrate comparative single-molecule studies of translation can now be done with genetically encoded tools alone. Finally, we use the HA frankenbody to precisely quantify the expression of HA-tagged proteins in developing zebrafish embryos. The versatility of the HA frankenbody makes it a powerful tool for imaging protein dynamics in vivo.

Suggested Citation

  • Ning Zhao & Kouta Kamijo & Philip D. Fox & Haruka Oda & Tatsuya Morisaki & Yuko Sato & Hiroshi Kimura & Timothy J. Stasevich, 2019. "A genetically encoded probe for imaging nascent and mature HA-tagged proteins in vivo," Nature Communications, Nature, vol. 10(1), pages 1-16, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-10846-1
    DOI: 10.1038/s41467-019-10846-1
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    Cited by:

    1. Daniel Strebinger & Chris J. Frangieh & Mirco J. Friedrich & Guilhem Faure & Rhiannon K. Macrae & Feng Zhang, 2023. "Cell type-specific delivery by modular envelope design," Nature Communications, Nature, vol. 14(1), pages 1-18, December.
    2. Malgorzata J. Latallo & Shaopeng Wang & Daoyuan Dong & Blake Nelson & Nathan M. Livingston & Rong Wu & Ning Zhao & Timothy J. Stasevich & Michael C. Bassik & Shuying Sun & Bin Wu, 2023. "Single-molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded RNA repeats in C9ORF72-ALS/FTD," Nature Communications, Nature, vol. 14(1), pages 1-18, December.
    3. Jiajun Xu & Ningning Zhu & Yijing Du & Tianyang Han & Xue Zheng & Jia Li & Shoujun Zhu, 2024. "Biomimetic NIR-II fluorescent proteins created from chemogenic protein-seeking dyes for multicolor deep-tissue bioimaging," Nature Communications, Nature, vol. 15(1), pages 1-16, December.
    4. Lance T. Denes & Chase P. Kelley & Eric T. Wang, 2021. "Microtubule-based transport is essential to distribute RNA and nascent protein in skeletal muscle," Nature Communications, Nature, vol. 12(1), pages 1-19, December.
    5. Charlotte A. Cialek & Gabriel Galindo & Tatsuya Morisaki & Ning Zhao & Taiowa A. Montgomery & Timothy J. Stasevich, 2022. "Imaging translational control by Argonaute with single-molecule resolution in live cells," Nature Communications, Nature, vol. 13(1), pages 1-14, December.

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