Abstract
The possibility to visualize small bacterial RNAs inside macrophages infected with Mycobacterium tuberculosis was demonstrated for the first time. A macrophage cell line was infected with the M. tuberculosis strain expressing small noncoding mycobacterial RNA MTS1338 fused with an RNA aptamer, which could bind a fluorophore and trigger its fluorescence. As a result, treatment of the infected macrophages with the DFHBI-1T fluorophore allowed fluorescence-based detection of the aptamer-labeled MTS1338 both in mycobacteria and in the host cell cytoplasm. This system can significantly aid in revealing the role of small M. tuberculosis RNAs in the pathogenesis of tuberculosis through identification of their secretion routes and eukaryotic targets and elucidation of the associated molecular pathways.
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ACKNOWLEDGMENTS
We are grateful to M.S. Baranov for kindly providing DFHBI-1T and N.G. Gurskaya for kindly providing the plasmids containing the Broccoli aptamers.
Funding
The study was supported by the Russian Science Foundation (project no. 18-15-00332).
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The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.
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Translated by M. Batrukova
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Bychenko, O.S., Skvortsova, Y.V., Grigorov, A.S. et al. Use of Genetically Encoded Fluorescent Aptamers for Visualization of Mycobacterium tuberculosis Small RNA MTS1338 in Infected Macrophages. Dokl Biochem Biophys 493, 185–189 (2020). https://doi.org/10.1134/S1607672920040055
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DOI: https://doi.org/10.1134/S1607672920040055