論文 - 詳細
| RRC ID | 59729 |
|---|---|
| 著者 | Farhana I, Hossain MN, Suzuki K, Matsuda T, Nagai T. |
| タイトル | Genetically Encoded Fluorescence/Bioluminescence Bimodal Indicators for Ca2+ Imaging. |
| ジャーナル | ACS Sens |
| Abstract |
Fluorescent and bioluminescent genetically encoded Ca2+ indicators (GECIs) are an indispensable tool for monitoring Ca2+ dynamics in numerous cellular events. Although fluorescent GECIs have a high spatiotemporal resolution, their application is often confined to short-term imaging due to the external illumination that causes phototoxicity and autofluorescence from specimens. Bioluminescent GECIs overcome these pitfalls with enhanced compatibility to optogenetic manipulation and photophysiological processes; however, they are compromised for spatiotemporal resolution. Therefore, there has been a push toward the use of Ca2+ indicators that possess the advantages of both fluorescent and bioluminescent GECI for a wide range of applications. To address this, we developed a high-affinity bimodal GECI, GLICO, using a single fluorescent protein-based GECI combined with a split luciferase. Through this novel design, the fusion protein becomes bimodal and possesses Ca2+ sensing properties similar to those of its fluorescent ancestor and confers bioluminescence-based Ca2+ imaging. GLICO in bioluminescence mode has the highest dynamic range (2200%) of all bioluminescent GECIs. We demonstrated the performance of GLICO in studying cytosolic Ca2+ dynamics in different cultured cells in each mode. With the purpose of Ca2+ imaging in high Ca2+ content organelle, we also created a low-affinity variant, ReBLICO and performed Ca2+ imaging of the ER in both fluorescence and bioluminescence modes. The ability to switch between fluorescence and bioluminescence modes with a single indicator would benefit transgenic applications by presenting an opportunity for a wide range of live Ca2+ imaging in physiological and pathophysiological conditions. |
| 巻・号 | 4(7) |
| ページ | 1825-1834 |
| 公開日 | 2019-7-26 |
| DOI | 10.1021/acssensors.9b00531 |
| PMID | 31276380 |
| MeSH | Animals Calcium / analysis* Calcium / metabolism Calmodulin / genetics Calmodulin / isolation & purification Calmodulin / metabolism* Cell Line, Tumor Channelrhodopsins / metabolism Fluorescence Green Fluorescent Proteins / genetics Green Fluorescent Proteins / isolation & purification Green Fluorescent Proteins / metabolism* Humans Neurons / metabolism Optical Imaging Protein Engineering Rats Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / isolation & purification Recombinant Fusion Proteins / metabolism* |
| IF | 6.944 |
| 引用数 | 5 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 25 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | HeLa |