Quantifying protein dynamics and stability in a living organism
Quantifying protein dynamics and stability in a living organism
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DOI：
10.1038/s41467-019-09088-y
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发表时间：
2019-03-12
影响因子：
16.6
通讯作者：
Davis, Caitlin M.
中科院分区：
文献类型：
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作者：
Feng, Ruopei;Gruebele, Martin;Davis, Caitlin M.
作为现代细胞生物学的一个组成部分，荧光显微镜能够量化培养细胞内荧光标记生物分子的稳定性和动态。然而，从活脊椎动物体内的单个细胞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管道，它将大范围核酸酶介导的嵌合转化与荧光检测的温度跳跃显微镜相结合，以探测活体多细胞生物不同组织中内源表达蛋白质的动态和稳定性。
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 temperaturejump microscopy to probe dynamics and stability of endogenously expressed proteins in different tissues of living multicellular organisms.