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科学家开发出探测1.95传奇私服发布网刚开一秒单细胞光学信号传导途径新方法

发布时间:2019-09-12 05:58

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本期文章:《自然—方法学》:Online/在线发表 日本京都大学Michiyuki Matsuda团队开发出在活体双光子(2P)光遗传学中,荧光共振能量转移(FRET)辅助的黄素蛋白的光活化技术。相关论文2019年9月9日在线发表在《自然方法学》上。 研究人员介绍,现有的光

期刊信息

附:英文原文

官方网址:https://www.nature.com/nmeth/

Nature Methods:《自然—方法学》,创刊于2004年。隶属于施普林格·自然出版集团,最新IF:28.467

Source:https://www.nature.com/articles/s41592-019-0541-5

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 Frster 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. Fusion to fluorescent proteins enables efficient two-photon activation of blue-light-controlled optical dimerizers via FRET. FRET-assisted photoactivation was used to study extracellular signal-regulated kinase activation in 3D epithelial cysts, organoids and living mice.

投稿链接:https://mts-nmeth.nature.com/cgi-bin/main.plex

Author: Tomoaki Kinjo, Kenta Terai, Shoichiro Horita, Norimichi Nomura, Kenta Sumiyama, Kaori Togashi, So Iwata, Michiyuki Matsuda

日本京都大学Michiyuki Matsuda团队开发出在活体双光子(2P)光遗传学中,荧光共振能量转移(FRET)辅助的黄素蛋白的光活化技术。相关论文2019年9月9日在线发表在《自然—方法学》上。

为了克服这一问题,研究人员开发了FRET辅助光活化技术,或称为FRAPA。在2P激发下,mTagBFP2有效地吸收能量并将能量转移到CRY2的发色团。基于结构引导工程,开发了具有40%FRET效率的嵌合蛋白,并命名为2P激活的CRY2或2paCRY2。2paCRY2用于开发名为2paRAF的RAF1激活系统。在表达2paRAF的三维培养细胞中,新开单职业传奇私服,细胞外信号调节激酶(ERK)在单细胞分辨率下通过2P激发被有效激活。在表达2paRAF的小鼠的表皮细胞中也完成了ERK的光激活。他们进一步开发了mTFP1融合的LOV结构域,其表现出对2P激发的有效响应。总的来说,FRAPA将为体内单细胞光学信号传导途径控制铺平道路。

Issue&Volume: 2019-09-09

研究人员介绍,现有的光学二聚体以优化信号传导为途径,但它们在体内的用途有限,部分原因是它们在2P激发下的低效活化。

DOI: 10.1038/s41592-019-0541-5

Title: FRET-assisted photoactivation of flavoproteins for in vivo two-photon optogenetics

本期文章:《自然—方法学》:Online/在线发表

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