論文 - 詳細
| RRC ID | 57649 |
|---|---|
| 著者 | Kinjo T, Terai K, Horita S, Nomura N, Sumiyama K, Togashi K, Iwata S, Matsuda M. |
| タイトル | FRET-assisted photoactivation of flavoproteins for in vivo two-photon optogenetics. |
| ジャーナル | Nat Methods |
| Abstract |
Optical dimerizers have been developed to untangle signaling pathways, but they are of limited use in vivo, partly due to their inefficient activation under two-photon (2P) excitation. To overcome this problem, we developed Förster resonance energy transfer (FRET)-assisted photoactivation, or FRAPA. On 2P excitation, mTagBFP2 efficiently absorbs and transfers the energy to the chromophore of CRY2. Based on structure-guided engineering, a chimeric protein with 40% FRET efficiency was developed and named 2P-activatable CRY2, or 2paCRY2. 2paCRY2 was employed to develop a RAF1 activation system named 2paRAF. In three-dimensionally cultured cells expressing 2paRAF, extracellular signal-regulated kinase (ERK) was efficiently activated by 2P excitation at single-cell resolution. Photoactivation of ERK was also accomplished in the epidermal cells of 2paRAF-expressing mice. We further developed an mTFP1-fused LOV domain that exhibits efficient response to 2P excitation. Collectively, FRAPA will pave the way to single-cell optical control of signaling pathways in vivo. |
| 巻・号 | 16(10) |
| ページ | 1029-1036 |
| 公開日 | 2019-10-1 |
| DOI | 10.1038/s41592-019-0541-5 |
| PII | 10.1038/s41592-019-0541-5 |
| PMID | 31501546 |
| MeSH | Animals Enzyme Activation Extracellular Signal-Regulated MAP Kinases / metabolism Flavoproteins / metabolism* Fluorescence Resonance Energy Transfer / methods* Mice Optogenetics* Photons* |
| IF | 30.822 |
| 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 | 42 |
| 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 | pCMV-VSV-G-RSV-Rev (RDB04393) |
| ヒト・動物細胞 | MDCK(RCB0995) |