Bright and stable monomeric green fluorescent protein derived from StayGold

Authors

Hanbin Zhang, Gleb D. Lesnov, Oksana M. Subach, Wenhao Zhang, Tatyana P. Kuzmicheva, Anna V. Vlaskina, Valeriya R. Samygina, Liangyi Chen, Xianxin Ye, Alena Yu. Nikolaeva, Azat Gabdulkhakov, Stavrini Papadaki, Wenming Qin, Valentin Borschevskiy, Maxim M. Perfilov, Alexey S. Gavrikov, Mikhail Drobizhev, Alexander S. Mishin, Kiryl D. Piatkevich, Fedor V. Subach

Publication

Nature Methods

Abstract

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.

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