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Bright and stable monomeric green fluorescent protein derived from StayGold

  • 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 Borshchevskiy,
  • Maxim M. Perfilov,
  • Alexey S. Gavrikov,
  • Mikhail Drobizhev,
  • Alexander S. Mishin,
  • Kiryl D. Piatkevich,
  • Fedor V. Subach

摘要

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 Caenorhabditiselegans and mice.