Optogenetic Control of Membrane Trafficking Using Light-Activated Reversible Inhibition by Assembly Trap of Intracellular Membranes (IM-LARIAT).

Bobae Hyeon, Mai Khanh Nguyen, Won Do Heo

Methods in molecular biology (Clifton, N.J.) 2022

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Intracellular membrane trafficking is a dynamic and complex cellular process. To study membrane trafficking with a high spatiotemporal resolution, we present an optogenetic method based on a blue-light inducible oligomerization of Rab GTPases, termed light-activated reversible inhibition by assembly trap of intracellular membranes (IM-LARIAT). In this chapter, we focus on the optical disruption of the dynamics and functions of previously studied intracellular membrane trafficking events, including transferrin recycling and growth cone regulation in relation to specific Rab GTPases. To aid general application, we provide a detailed description of transfection, imaging with a confocal microscope, and analysis of data. © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Bobae Hyeon, Mai Khanh Nguyen, Won Do Heo. Optogenetic Control of Membrane Trafficking Using Light-Activated Reversible Inhibition by Assembly Trap of Intracellular Membranes (IM-LARIAT). Methods in molecular biology (Clifton, N.J.). 2022;2473:309-331