論文 - 詳細
| RRC ID | 81679 |
|---|---|
| 著者 | Hashizume R, Fujii H, Mehta S, Ota K, Qian Y, Zhu W, Drobizhev M, Nasu Y, Zhang J, Bito H, Campbell RE. |
| タイトル | A genetically encoded far-red fluorescent calcium ion biosensor derived from a biliverdin-binding protein. |
| ジャーナル | Protein Sci |
| Abstract |
Far-red and near-infrared (NIR) genetically encoded calcium ion (Ca2+ ) indicators (GECIs) are powerful tools for in vivo and multiplexed imaging of neural activity and cell signaling. Inspired by a previous report to engineer a far-red fluorescent protein (FP) from a biliverdin (BV)-binding NIR FP, we have developed a far-red fluorescent GECI, designated iBB-GECO1, from a previously reported NIR GECI. iBB-GECO1 exhibits a relatively high molecular brightness, an inverse response to Ca2+ with ΔF/Fmin = -13, and a near-optimal dissociation constant (Kd ) for Ca2+ of 105 nM. We demonstrate the utility of iBB-GECO1 for four-color multiplexed imaging in MIN6 cells and five-color imaging in HEK293T cells. Like other BV-binding GECIs, iBB-GECO1 did not give robust signals during in vivo imaging of neural activity in mice, but did provide promising results that will guide future engineering efforts. SIGNIFICANCE: Genetically encoded calcium ion (Ca2+ ) indicators (GECIs) compatible with common far-red laser lines (~630-640 nm) on commercial microscopes are of critical importance for their widespread application to deep-tissue multiplexed imaging of neural activity. In this study, we engineered a far-red excitable fluorescent GECI, designated iBB-GECO1, that exhibits a range of preferable specifications such as high brightness, large fluorescence response to Ca2+ , and compatibility with multiplexed imaging in mammalian cells. |
| 巻・号 | 31(10) |
| ページ | e4440 |
| 公開日 | 2022-10-1 |
| DOI | 10.1002/pro.4440 |
| PMID | 36173169 |
| PMC | PMC9518226 |
| MeSH | Animals Biliverdine* / metabolism Biosensing Techniques* Calcium / metabolism Carrier Proteins HEK293 Cells Humans Ions Luminescent Proteins / chemistry Mice |
| IF | 3.876 |
| 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 | 5 |
| 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 | |
| 遺伝子材料 | pCAG-iBB-GECO1 (RDB20303) pCaMKII-iBB-GECO1 (RDB20304) pCAGGS (RDB08938) |