Video/Audio Open Access

Magnetic manipulation of intracellular signals

Elie BALLOUL

Other(s)
John Manzi; Fanny Cayrac
Project member(s)
Koceila Aizel; Cornelia Monzel; Maud Bongaerts
Supervisor(s)
Mathieu Coppey; Maxime Dahan

In the context of the MAGNEURON project we have developed magnetic tools to manipulate intracellular proteins involved in intracellular signaling processes. We focused on the manipulation of the subdomain DHPH of ITSN1 in order to control the activity of CDC42.

To control the activity of CDC42 we produced a fusion protein consisting in the DHPH domain of ITSN1, the mCherry fluorescent protein and a nanobody against GFP (aGFPnb).

The manipulations were done either using a magnetic tip or magnetic micropillars done with soft magnetic material (Nickel Iron alloy). Nanoparticles were first injected in the cytoplasm of the cell. After a delay of at least ten minutes, the survival of the cell was controlled and the manipulation could start.
We monitored the fluorescence of the reporter (IRFP-NWASP) and the formation of protrusion/filopodia as markers of a biological response to the manipulation.

We observed protrusion formation as well as NWASP activity when attracting the nanoparticles, but control experiments (Manipulation of particles without ITSN1-mcherry-aGFPnb) also showed increase of fluorescence of the NWASP reporter at the point of attraction of the particles.

In addition, if compared to optogenetic manipulation of the similar ITSN1-DHPH domain, the formation of protrusion was very limited. We concluded that magnetic manipulation of intracellular signals was not efficient to hijack signaling pathways.

We publish here the raw data of some experiments that were produced in the lab.

Each experiment has a README file describing the parameter of the recording and what can be observe on the video

Files (1.7 GB)
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manip032018_fMNPs_ITSN1_NWASP_Hela_cells.zip
md5:8027e007fb2d7032a91a9d8e43276bc3
236.8 MB Download
manip052018_fMNPs_ITSN1_NWASP_Hela_cells.zip
md5:2bb2ae012183eaf2d66c8a591ccdab97
177.8 MB Download
manip062018_fMNPs_ITSN1_NWASP_Hela_cells.zip
md5:396080d16feaab760487d8a03d5aecb1
255.4 MB Download
manip092019_control.zip
md5:ff877641e9c7c964abd93b6165b7ff5a
956.7 MB Download
manip102017_fMNPs_ITSN1_NWASP_Hela_cells.zip
md5:f522e9b3b047fca926ae95dd37e9165a
115.1 MB Download
  • Toraille, L., Aïzel, K., Balloul, É., Vicario, C., Monzel, C., Coppey, M., Secret, E., Siaugue, J.-M., Sampaio, J., Rohart, S., Vernier, N., Bonnemay, L., Debuisschert, T., Rondin, L., Roch, J.-F., Dahan, M., 2018. Optical Magnetometry of Single Biocompatible Micromagnets for Quantitative Magnetogenetic and Magnetomechanical Assays. Nano Letters. https://doi.org/10.1021/acs.nanolett.8b03222

  • Monzel, C., Vicario, C., Piehler, J., Coppey, M., Dahan, M., 2017. Magnetic control of cellular processes using biofunctional nanoparticles. Chemical Science 8, 7330–7338. https://doi.org/10.1039/C7SC01462G

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