Agostino, VS, Trinconi, CM, Galuppo, MK, Price, HP ORCID: https://orcid.org/0000-0003-1537-4390 and Uliana, SRB (2020) Evaluation of NanoLuc, RedLuc and Luc2 as bioluminescent reporters in a cutaneous leishmaniasis model. Acta Tropica. 105444 - 105444.

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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.

Item Type: Article
Additional Information: The final accepted version of this article and all relevant information can be found at; https://www.sciencedirect.com/science/article/abs/pii/S0001706X20300097?via%3Dihub
Uncontrolled Keywords: Leishmania; Drug screening; Bioluminescent reporters; Bioimaging; NanoLuc; Red-shifted luciferase
Subjects: Q Science > Q Science (General)
Q Science > QR Microbiology
Divisions: Faculty of Natural Sciences > School of Life Sciences
Depositing User: Symplectic
Date Deposited: 19 Mar 2020 15:20
Last Modified: 19 Mar 2020 15:20
URI: http://eprints.keele.ac.uk/id/eprint/7815

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