School of Life Sciences, Westlake University, Hangzhou, China.
Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, China.
Nat Methods. 2024 Apr;21(4):657-665. doi: 10.1038/s41592-024-02203-y. Epub 2024 Feb 26.
The high brightness and photostability of the green fluorescent protein StayGold make it a particularly attractive probe for long-term live-cell imaging; however, its dimeric nature precludes its application as a fluorescent tag for some proteins. Here, we report the development and crystal structures of a monomeric variant of StayGold, named mBaoJin, which preserves the beneficial properties of its precursor, while serving as a tag for structural proteins and membranes. Systematic benchmarking of mBaoJin against popular green fluorescent proteins and other recently introduced monomeric and pseudomonomeric derivatives of StayGold established mBaoJin as a bright and photostable fluorescent protein, exhibiting rapid maturation and high pH/chemical stability. mBaoJin was also demonstrated for super-resolution, long-term live-cell imaging and expansion microscopy. We further showed the applicability of mBaoJin for neuronal labeling in model organisms, including Caenorhabditis elegans and mice.
绿色荧光蛋白 StayGold 的高亮度和光稳定性使其成为长期活细胞成像的特别有吸引力的探针;然而,其二聚体性质排除了其作为某些蛋白质的荧光标记的应用。在这里,我们报告了 StayGold 的单体变体 mBaoJin 的开发和晶体结构,该变体保留了其前体的有益特性,同时作为结构蛋白和膜的标记。mBaoJin 与流行的绿色荧光蛋白以及其他最近引入的 StayGold 的单体和拟单体衍生物的系统基准测试确立了 mBaoJin 作为一种明亮且稳定的荧光蛋白,表现出快速成熟和高 pH/化学稳定性。mBaoJin 还可用于超分辨率、长期活细胞成像和扩展显微镜。我们还进一步展示了 mBaoJin 在包括秀丽隐杆线虫和小鼠在内的模式生物中的神经元标记中的适用性。