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Proximity-dependent initiation of hybridization chain reaction

Author

Listed:
  • Björn Koos

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Gaëlle Cane

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Karin Grannas

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Liza Löf

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Linda Arngården

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Johan Heldin

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Carl-Magnus Clausson

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Axel Klaesson

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • M. Karoliina Hirvonen

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Felipe M. S. de Oliveira

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Vladimir O. Talibov

    (Box 256, Uppsala University)

  • Nhan T. Pham

    (School of Biological Sciences and School of Biomedical Sciences, University of Edinburgh, C H Waddington Building, Max Born Cresent, Kings Buildings, Edinburgh EH9 3BF, UK)

  • Manfred Auer

    (School of Biological Sciences and School of Biomedical Sciences, University of Edinburgh, C H Waddington Building, Max Born Cresent, Kings Buildings, Edinburgh EH9 3BF, UK)

  • U. Helena Danielson

    (Box 256, Uppsala University)

  • Johannes Haybaeck

    (Institute of Pathology, Medical University of Graz)

  • Masood Kamali-Moghaddam

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

  • Ola Söderberg

    (Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center)

Abstract

Sensitive detection of protein interactions and post-translational modifications of native proteins is a challenge for research and diagnostic purposes. A method for this, which could be used in point-of-care devices and high-throughput screening, should be reliable, cost effective and robust. To achieve this, here we design a method (proxHCR) that combines the need for proximal binding with hybridization chain reaction (HCR) for signal amplification. When two oligonucleotide hairpins conjugated to antibodies bind in close proximity, they can be activated to reveal an initiator sequence. This starts a chain reaction of hybridization events between a pair of fluorophore-labelled oligonucleotide hairpins, generating a fluorescent product. In conclusion, we show the applicability of the proxHCR method for the detection of protein interactions and posttranslational modifications in microscopy and flow cytometry. As no enzymes are needed, proxHCR may be an inexpensive and robust alternative to proximity ligation assays.

Suggested Citation

  • Björn Koos & Gaëlle Cane & Karin Grannas & Liza Löf & Linda Arngården & Johan Heldin & Carl-Magnus Clausson & Axel Klaesson & M. Karoliina Hirvonen & Felipe M. S. de Oliveira & Vladimir O. Talibov & N, 2015. "Proximity-dependent initiation of hybridization chain reaction," Nature Communications, Nature, vol. 6(1), pages 1-10, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8294
    DOI: 10.1038/ncomms8294
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    Cited by:

    1. Doroteya Raykova & Despoina Kermpatsou & Tony Malmqvist & Philip J. Harrison & Marie Rubin Sander & Christiane Stiller & Johan Heldin & Mattias Leino & Sara Ricardo & Anna Klemm & Leonor David & Ola S, 2022. "A method for Boolean analysis of protein interactions at a molecular level," Nature Communications, Nature, vol. 13(1), pages 1-17, December.
    2. Sungwook Woo & Sinem K. Saka & Feng Xuan & Peng Yin, 2024. "Molecular robotic agents that survey molecular landscapes for information retrieval," Nature Communications, Nature, vol. 15(1), pages 1-12, December.

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