DMN-Tre Labeling for Detection and High-Content Screening of Compounds against Intracellular Mycobacteria

  • ACS Omega. 2020 Feb 14;5(7):3661-3669. doi: 10.1021/acsomega.9b04173.
Henok A Sahile  1 ,  Celine Rens  1 ,  Tirosh Shapira  1 ,  Raymond J Andersen  2 ,  Yossef Av-Gay  1  1
Affiliations
  • 1. Division of Infectious Diseases, Department of Medicine and Department of Microbiology and Immunology, Life Sciences Institute, University of British Columbia, 2350 Health Sciences Mall, Vancouver, British Columbia, Canada V6T 1Z3.
  • 2. Department of Earth, Ocean and Atmospheric Sciences, Faculty of Science, University of British Columbia, 2036 Main Mall, Vancouver, British Columbia, Canada V6T 1Z1.
PMID: 32118181 DOI: 10.1021/acsomega.9b04173
Abstract

4-N,N-Dimethylamino-1,8-naphthalimide conjugate of trehalose (DMN-Tre) is a fluorogenic dye recently developed as a diagnostic tool for tuberculosis. DMN-Tre selectively labels the mycobacterial cell wall through the Ag85 Enzymes. In this work, we disclose a protocol describing the total synthesis of DMN-Tre with more than 99% purity. We further developed a protocol for in vitro and intercellular labeling of various mycobacterial strains. DMN-Tre labeling was found to be a useful tool to study in vitro and intracellular Mycobacterium tuberculosis (Mtb) physiology and as an end-point readout system in high-content image-based screening (HCS) of drug molecules. Such uses of DMN-Tre labeling provide a simple, fast, and cheap alternative to the existing, time-consuming approach that requires Mtb strains to be genetically transformed with fluorescent reporter genes.

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