Red light-emitting short Mango-based system enables tracking a mycobacterial small noncoding RNA in infected macrophages.
Oksana S BychenkoAlexei A KhrulevJulia I SvetlovaVladimir B TsvetkovPolina N KamzeevaYulia V SkvortsovaBoris S TupertsevIgor A IvanovLeonid V AseevYuriy M KhodarovichEvgeny S BelyaevLiubov I KozlovskayaTimofei S ZatsepinTatyana L AzhikinaAnna M VarizhukMikhail S BaranovPublished in: Nucleic acids research (2023)
Progress in RNA metabolism and function studies relies largely on molecular imaging systems, including those comprising a fluorogenic dye and an aptamer-based fluorescence-activating tag. G4 aptamers of the Mango family, typically combined with a duplex/hairpin scaffold, activate the fluorescence of a green light-emitting dye TO1-biotin and hold great promise for intracellular RNA tracking. Here, we report a new Mango-based imaging platform. Its key advantages are the tunability of spectral properties and applicability for visualization of small RNA molecules that require minimal tag size. The former advantage is due to an expanded (green-to-red-emitting) palette of TO1-inspired fluorogenic dyes, and the truncated duplex scaffold ensures the latter. To illustrate the applicability of the improved platform, we tagged Mycobacterium tuberculosis sncRNA with the shortened aptamer-scaffold tag. Then, we visualized it in bacteria and bacteria-infected macrophages using the new red light-emitting Mango-activated dye.
Keyphrases
- light emitting
- mycobacterium tuberculosis
- nucleic acid
- gold nanoparticles
- tissue engineering
- highly efficient
- high throughput
- single molecule
- signaling pathway
- sensitive detection
- high resolution
- aqueous solution
- optical coherence tomography
- machine learning
- magnetic resonance
- magnetic nanoparticles
- reactive oxygen species
- photodynamic therapy
- deep learning
- fluorescence imaging
- fluorescent probe