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Evaluation of NanoLuc, RedLuc and Luc2 as bioluminescent reporters in a cutaneous leishmaniasis model. | LitMetric

Evaluation of NanoLuc, RedLuc and Luc2 as bioluminescent reporters in a cutaneous leishmaniasis model.

Acta Trop

Department of Parasitology, Biomedical Sciences Institute, University of Sao Paulo, Sao Paulo CEP 05508-000, Brazil. Electronic address:

Published: June 2020

AI Article Synopsis

  • Researchers have developed three new Leishmania amazonensis mutants that express various luciferases to improve drug screening efficiency.
  • While NanoLuc shows high light intensity in lab tests, it fails to distinguish between live and dead parasites; RedLuc and Luc2 provide more reliable results in both lab and animal studies for evaluating drug effectiveness.

Article Abstract

New drugs for the treatment of human leishmaniasis are urgently needed, considering the limitations of current available options. However, pre-clinical evaluation of drug candidates for leishmaniasis is challenging. The use of luciferase-expressing parasites for parasite load detection is a potentially powerful tool to accelerate the drug discovery process. We have previously described the use of Leishmania amazonensis mutants expressing firefly luciferase (Luc2) for drug testing. Here, we describe three new mutant L. amazonensis lines that express different variants of luciferases: NanoLuc, NanoLuc-PEST and RedLuc. These mutants were evaluated in drug screening protocols. NanoLuc-parasites, in spite of high bioluminescence intensity in vitro, were shown to be inadequate in discriminating between live and dead parasites. Bioluminescence detection from intracellular amastigotes expressing NanoLuc-PEST, RedLuc or Luc2 proved more reliable than microscopy to determine parasite killing. Increased sensitivity was observed in vivo with RedLuc-expressing parasites as compared to NanoLuc-expressing L. amazonensis. Our data indicates that NanoLuc is not suitable for in vivo parasite burden determination. Additionally, RedLuc and the conventional luciferase Luc2 demonstrated equivalent sensitivity in an in vivo model of cutaneous leishmaniasis.

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Source
http://dx.doi.org/10.1016/j.actatropica.2020.105444DOI Listing

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