Video/Audio Open Access
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
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