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Comparison of monomeric variants of StayGold

Miyawaki, A.; Shimozono, S.; Ando, R.; Sugiyama, M.; Hirano, M.; Niino, Y.

2024-02-29 bioengineering
10.1101/2024.02.28.582207 bioRxiv
Show abstract

StayGold is a bright and highly photostable fluorescent protein (FP) that forms an obligate dimer, thereby limiting its application as a soluble marker. On the basis of the structural information of this FP, we disrupted the dimerization to generate a monomeric variant, mStayGold, which inherits both the extremely high photostability and the high practical brightness of StayGold, for molecular fusion and membrane-targeting applications. Meanwhile, two other research groups have independently monomerized StayGold using different strategies. As a result, multiple StayGold monomers are currently available, creating confusion in the research community. In the present study, we investigated the three basic properties--photostability, brightness, and dispersibility--of these monomers by performing detailed side-by-side comparisons. This study highlights the difficulties of StayGold monomerization with emphasis on the tradeoff between photostability and brightness in FP technology.

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