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Tohoku University Technology: Fluorescent Dyes for Detection of Live Cell Nucleolar RNA: T20-457 and others

Achieves high brightness, high light stability, and clear light-up response; can also be applied to envelope virus detection and vaccine quality testing.

There is only one commercially available intracellular nucleolar RNA dye, SYTO RNASelect. This dye has challenges such as being unsuitable for live cells, having poor photostability, and emitting fluorescence at relatively short wavelengths. The BIOP of the present invention is a novel RNA dye that shows a fluorescence quantum yield of 0.42 when bound to RNA, enabling high-brightness imaging of RNA in live cells. It also has excellent cell membrane permeability, allowing for high-contrast staining of nucleolar RNA in live cells in a short time. Furthermore, it shows almost no cytotoxicity, allowing for long-term observation. Due to its excellent fluorescent properties and membrane permeability, BIOP can also be applied for the rapid detection (~10 minutes) of enveloped viruses containing genomic RNA. Additionally, BIOP is useful for quality control of mRNA vaccines. It has been found to be applicable for measuring the RNA encapsulation rate of vaccines and RNA leakage based on vaccine quality degradation. When using a model formulation of mRNA vaccines, the detection limit of BIOP is approximately 10^7 particles/mL, which is considered sufficiently sensitive for vaccines like Pfizer's BNT162b2 (approximately 4×10^13 particles/mL).

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