Related Experiment Video
Updated: Dec 9, 2025
A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening
Published on: January 17, 2014
Use of Genetically Encoded Fluorescent Aptamers for Visualization of Mycobacterium tuberculosis Small RNA MTS1338 in
O S Bychenko1, Yu V Skvortsova2, A S Grigorov2
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia. bychenko.oksana@gmail.com.
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.
