Shiori Takeuchi, Shosei Imai, Takuya Terai and Robert E. Campbell
{"title":"A chemigenetic indicator based on a synthetic chelator and a green fluorescent protein for imaging of intracellular sodium ions†","authors":"Shiori Takeuchi, Shosei Imai, Takuya Terai and Robert E. Campbell","doi":"10.1039/D4CB00256C","DOIUrl":null,"url":null,"abstract":"<p >A chemigenetic indicator with an affinity suitable for imaging of intracellular sodium ions (Na<small><sup>+</sup></small>) 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<small><sup>+</sup></small> and an ionophore.</p>","PeriodicalId":40691,"journal":{"name":"RSC Chemical Biology","volume":" 2","pages":" 170-174"},"PeriodicalIF":3.1000,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11638762/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"RSC Chemical Biology","FirstCategoryId":"1085","ListUrlMain":"https://pubs.rsc.org/en/content/articlelanding/2025/cb/d4cb00256c","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
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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.
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