Optimized iLID Membrane Anchors for Local Optogenetic Protein Recruitment

• DOI: 10.1021/acssynbio.0c00511
• Publication Date: 2021-04-16T00:33:30Z
• Authors: Dean E. Natwick; Sean R. Collins
• Keywords: Component (thermodynamics)

Optogenetic protein dimerization systems are powerful tools to investigate the biochemical networks that cells use make decisions and coordinate their activities. These tools, including improved Light-Inducible Dimer (iLID) system, offer ability selectively recruit components subcellular locations, such as micron-scale regions of plasma membrane. In this way, role individual proteins within signaling can be examined with high spatiotemporal resolution. Currently, consistent recruitment is limited by heterogeneous optogenetic component expression, spatial precision diminished diffusion, especially over long time scales. Here, we address these challenges iLID system alternative membrane anchoring domains fusion configurations. Using live cell imaging mathematical modeling, demonstrate strategy affects both expression which in turn impact strength, kinetics, dynamics. Compared commonly used C-terminal fusion, large N-terminal anchors show stronger local recruitment, slower diffusion recruited components, efficient wider gene ranges, control outputs. We also define guidelines for regimes optimal cell-wide strategies. Our findings highlight key sources imprecision light-inducible dimer provide allow greater localization across diverse biological applications.