定量研究活生物体中的蛋白质动态和稳定性。
Quantifying protein dynamics and stability in a living organism.
机构信息
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
出版信息
Nat Commun. 2019 Mar 12;10(1):1179. doi: 10.1038/s41467-019-09088-y.
As an integral part of modern cell biology, fluorescence microscopy enables quantification of the stability and dynamics of fluorescence-labeled biomolecules inside cultured cells. However, obtaining time-resolved data from individual cells within a live vertebrate organism remains challenging. Here we demonstrate a customized pipeline that integrates meganuclease-mediated mosaic transformation with fluorescence-detected temperature-jump microscopy to probe dynamics and stability of endogenously expressed proteins in different tissues of living multicellular organisms.
作为现代细胞生物学的一个组成部分，荧光显微镜使我们能够定量测量培养细胞内荧光标记生物分子的稳定性和动态变化。然而，要从活体脊椎动物体内的单个细胞中获取时间分辨数据仍然具有挑战性。在这里，我们展示了一个定制的流水线，该流水线将 meganuclease 介导的马赛克转化与荧光检测的温度跃变显微镜相结合，用于探测活体多细胞生物不同组织中内源性表达蛋白的动力学和稳定性。
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