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Versatile protein tagging in cells with split fluorescent protein

Author

Listed:
  • Daichi Kamiyama

    (University of California)

  • Sayaka Sekine

    (University of California)

  • Benjamin Barsi-Rhyne

    (Tetrad Graduate Program, University of California)

  • Jeffrey Hu

    (University of California)

  • Baohui Chen

    (University of California)

  • Luke A. Gilbert

    (University of California)

  • Hiroaki Ishikawa

    (University of California)

  • Manuel D. Leonetti

    (University of California)

  • Wallace F. Marshall

    (University of California)

  • Jonathan S. Weissman

    (University of California
    Howard Hughes Medical Institute)

  • Bo Huang

    (University of California
    University of California)

Abstract

In addition to the popular method of fluorescent protein fusion, live cell protein imaging has now seen more and more application of epitope tags. The small size of these tags may reduce functional perturbation and enable signal amplification. To address their background issue, we adapt self-complementing split fluorescent proteins as epitope tags for live cell protein labelling. The two tags, GFP11 and sfCherry11 are derived from the eleventh β-strand of super-folder GFP and sfCherry, respectively. The small size of FP11-tags enables a cost-effective and scalable way to insert them into endogenous genomic loci via CRISPR-mediated homology-directed repair. Tandem arrangement FP11-tags allows proportional enhancement of fluorescence signal in tracking intraflagellar transport particles, or reduction of photobleaching for live microtubule imaging. Finally, we show the utility of tandem GFP11-tag in scaffolding protein oligomerization. These experiments illustrate the versatility of FP11-tag as a labelling tool as well as a multimerization-control tool for both imaging and non-imaging applications.

Suggested Citation

  • Daichi Kamiyama & Sayaka Sekine & Benjamin Barsi-Rhyne & Jeffrey Hu & Baohui Chen & Luke A. Gilbert & Hiroaki Ishikawa & Manuel D. Leonetti & Wallace F. Marshall & Jonathan S. Weissman & Bo Huang, 2016. "Versatile protein tagging in cells with split fluorescent protein," Nature Communications, Nature, vol. 7(1), pages 1-9, April.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms11046
    DOI: 10.1038/ncomms11046
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    1. Takeshi Hori & Hiroaki Okae & Shun Shibata & Norio Kobayashi & Eri H. Kobayashi & Akira Oike & Asato Sekiya & Takahiro Arima & Hirokazu Kaji, 2024. "Trophoblast stem cell-based organoid models of the human placental barrier," Nature Communications, Nature, vol. 15(1), pages 1-15, December.
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    5. Lee B. Miles & Vanessa Calcinotto & Sara Oveissi & Rita J. Serrano & Carmen Sonntag & Orlen Mulia & Clara Lee & Robert J. Bryson-Richardson, 2024. "CRIMP: a CRISPR/Cas9 insertional mutagenesis protocol and toolkit," Nature Communications, Nature, vol. 15(1), pages 1-11, December.
    6. Calvin Dumesnil & Lauri Vanharanta & Xavier Prasanna & Mohyeddine Omrane & Maxime Carpentier & Apoorva Bhapkar & Giray Enkavi & Veijo T. Salo & Ilpo Vattulainen & Elina Ikonen & Abdou Rachid Thiam, 2023. "Cholesterol esters form supercooled lipid droplets whose nucleation is facilitated by triacylglycerols," Nature Communications, Nature, vol. 14(1), pages 1-13, December.
    7. Sayaka Sekine & Mitsusuke Tarama & Housei Wada & Mustafa M. Sami & Tatsuo Shibata & Shigeo Hayashi, 2024. "Emergence of periodic circumferential actin cables from the anisotropic fusion of actin nanoclusters during tubulogenesis," Nature Communications, Nature, vol. 15(1), pages 1-13, December.

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