Han Kai, Huang Shuhan, Kong Jie, Yang Yang, Shi Lei, Ci Yali
State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing, China.
Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing, China.
FEBS Lett. 2023 Mar;597(5):693-701. doi: 10.1002/1873-3468.14581. Epub 2023 Feb 1.
Endoplasmic reticulum (ER) is a highly complicated and dynamic organelle that actively changes its shape and communicates with other organelles. Visualization of ER in live cells is of great importance to understand cellular activities. Here, we designed a novel ER marker, RR-mNeonGreen, which comprised an N-terminal ER retention signal, a bright fluorescent protein (mNeonGreen), and a C-terminal transmembrane region. Colocalization of RR-mNeonGreen with mCherry-KDEL verified that RR-mNeonGreen perfectly labeled the ER. RR-mNeonGreen showed better continuity of ER tubules when imaged by super-resolution microscopy. Moreover, RR-mNeonGreen is competent for live-cell imaging of ER dynamics and tracing of the interaction between ER and mitochondria at high spatiotemporal resolution. In summary, RR-mNeonGreen is a novel ER marker for super-resolution live-cell imaging with multiple merits.
内质网（ER）是一种高度复杂且动态的细胞器，它会积极改变其形状并与其他细胞器进行通讯。在活细胞中对内质网进行可视化对于理解细胞活动非常重要。在此，我们设计了一种新型的内质网标记物RR-mNeonGreen，它由一个N端内质网滞留信号、一种明亮的荧光蛋白（mNeonGreen）和一个C端跨膜区域组成。RR-mNeonGreen与mCherry-KDEL的共定位证实RR-mNeonGreen完美地标记了内质网。当通过超分辨率显微镜成像时，RR-mNeonGreen显示出更好的内质网管连续性。此外，RR-mNeonGreen能够以高时空分辨率对内质网动态进行活细胞成像以及追踪内质网与线粒体之间的相互作用。总之，RR-mNeonGreen是一种具有多种优点的用于超分辨率活细胞成像的新型内质网标记物。
