A chemigenetic indicator based on a synthetic chelator and a green fluorescent protein for imaging of intracellular sodium ions

Shiori Takeuchi; Shosei Imai; Takuya Terai; Robert E Campbell

doi:10.1039/d4cb00256c

A chemigenetic indicator based on a synthetic chelator and a green fluorescent protein for imaging of intracellular sodium ions

RSC Chem Biol. 2024 Dec 3. doi: 10.1039/d4cb00256c. Online ahead of print.

Authors

Shiori Takeuchi¹, Shosei Imai¹, Takuya Terai¹, Robert E Campbell^{1

2}

Affiliations

¹ Department of Chemistry, Graduate School of Science, The University of Tokyo 7-3-1 Hongo Bunkyo-ku Tokyo 113-0033 Japan terai@chem.s.u-tokyo.ac.jp campbell@chem.s.u-tokyo.ac.jp.
² CERVO Brain Research Center and Department of Biochemistry, Microbiology, and Bioinformatics, Université Laval, Québec Québec G1V 0A6 Canada.

Abstract

A chemigenetic indicator with an affinity suitable for imaging of intracellular sodium ions (Na⁺) in mammalian cells was developed. The indicator, based on a chimera of green fluorescent protein (GFP) and HaloTag labeled with a synthetic crown ether chelator, was produced by a combination of rational design and directed evolution. In mammalian cells the indicator exhibited an approximately 100% increase in excitation ratio when the cells were treated with 20 mM Na⁺ and an ionophore.