Purba Kashyap, Sara Bertelli, Fakun Cao, Yulia Kostritskaia, Fenja Blank, Niranjan A Srikanth, Claire Schlack-Leigers, Roberto Saleppico, Dolf Bierhuizen, Xiaocen Lu, Walter Nickel, Robert E Campbell, Andrew J R Plested, Tobias Stauber, Marcus J Taylor, Helge Ewers
We developed a system for optogenetic release of single molecules in cells. We confined soluble and transmembrane proteins to the Golgi apparatus via a photocleavable protein and released them by short pulses of light. Our method allows for a light dose-dependent delivery of functional proteins to the cytosol and plasma membrane in amounts compatible with single-molecule imaging, greatly simplifying access to single-molecule microscopy of any protein in live cells. We were able to reconstitute ion conductance by delivering BK and LRRC8/volume-regulated anion channels to the plasma membrane...
March 8, 2024: Nature Methods