小柯机器人

日本理化学研究所Atsushi Miyawaki、北里大学Kazuhiko Katayama等研究人员合作开发出一种高度光稳定的亮绿色荧光蛋白。该项研究成果于2022年4月25日在线发表在《自然—生物技术》杂志上。

研究人员提出了StayGold，一种来自水母Cytaeis uchidae的绿色荧光蛋白（GFP）。StayGold的光稳定性比目前任何可用的荧光蛋白高一个数量级，并且具有与mNeonGreen相似的细胞亮度。研究人员使用StayGold在几分钟内用结构照明显微镜（SIM）对内质网（ER）的动态进行了高时空分辨率成像，并观察到比GFP变体在ER中的稳定性要小得多的光漂白。

利用StayGold融合和SIM，研究人员还对线粒体融合和分裂的动态进行了成像，并绘制了感染严重急性呼吸道综合征冠状病毒2的固定细胞中的病毒突刺蛋白。由于StayGold是一个二聚体，研究人员创建了一个串联二聚体版本，使其能够观察微管的动态和神经元的兴奋性突触后密度。StayGold将大大减少光漂白带来的限制，特别是在活体细胞或体积成像中。

据悉，荧光蛋白的低光稳定性是许多荧光显微镜应用中的一个限制因素。

附：英文原文

Title: A highly photostable and bright green fluorescent protein

Author: Hirano, Masahiko, Ando, Ryoko, Shimozono, Satoshi, Sugiyama, Mayu, Takeda, Noriyo, Kurokawa, Hiroshi, Deguchi, Ryusaku, Endo, Kazuki, Haga, Kei, Takai-Todaka, Reiko, Inaura, Shunsuke, Matsumura, Yuta, Hama, Hiroshi, Okada, Yasushi, Fujiwara, Takahiro, Morimoto, Takuya, Katayama, Kazuhiko, Miyawaki, Atsushi

Issue&Volume: 2022-04-25

Abstract: The low photostability of fluorescent proteins is a limiting factor in many applications of fluorescence microscopy. Here we present StayGold, a green fluorescent protein (GFP) derived from the jellyfish Cytaeis uchidae. StayGold is over one order of magnitude more photostable than any currently available fluorescent protein and has a cellular brightness similar to mNeonGreen. We used StayGold to image the dynamics of the endoplasmic reticulum (ER) with high spatiotemporal resolution over several minutes using structured illumination microscopy (SIM) and observed substantially less photobleaching than with a GFP variant optimized for stability in the ER. Using StayGold fusions and SIM, we also imaged the dynamics of mitochondrial fusion and fission and mapped the viral spike proteins in fixed cells infected with severe acute respiratory syndrome coronavirus 2. As StayGold is a dimer, we created a tandem dimer version that allowed us to observe the dynamics of microtubules and the excitatory post-synaptic density in neurons. StayGold will substantially reduce the limitations imposed by photobleaching, especially in live cell or volumetric imaging. StayGold is over one order of magnitude more photostable than current fluorescent proteins

DOI: 10.1038/s41587-022-01278-2

Source: https://www.nature.com/articles/s41587-022-01278-2

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex