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细胞内光遗传学捕获导致的蛋白质可逆失活。

Reversible protein inactivation by optogenetic trapping in cells.

机构信息

1] Center for Cognition and Sociality, Institute for Basic Science (IBS), Daejeon, Republic of Korea. [2] Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea. [3].

1] Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea. [2].

出版信息

Nat Methods. 2014 Jun;11(6):633-6. doi: 10.1038/nmeth.2940. Epub 2014 May 4.

Abstract

We present a versatile platform to inactivate proteins in living cells using light, light-activated reversible inhibition by assembled trap (LARIAT), which sequesters target proteins into complexes formed by multimeric proteins and a blue light-mediated heterodimerization module. Using LARIAT, we inhibited diverse proteins that modulate cytoskeleton, lipid signaling and cell cycle with high spatiotemporal resolution. Use of single-domain antibodies extends the method to target proteins containing specific epitopes, including GFP.

摘要

我们提出了一种通用的平台,利用光来使活细胞中的蛋白质失活,即光激活的可逆抑制物组装陷阱(LARIAT),它将靶蛋白隔离到由多聚体蛋白和蓝光介导的异二聚化模块形成的复合物中。利用 LARIAT,我们以高时空分辨率抑制了多种调节细胞骨架、脂质信号和细胞周期的蛋白质。使用单域抗体将该方法扩展到针对含有特定表位的蛋白质,包括 GFP。

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