Skip to main content

Research Repository

Advanced Search

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

Agostino, VS; Trinconi, CM; Galuppo, MK; Price, H; Uliana, SRB

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


Authors

VS Agostino

CM Trinconi

MK Galuppo

SRB Uliana



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.

Journal Article Type Article
Acceptance Date Mar 11, 2020
Publication Date Mar 13, 2020
Journal Acta Tropica
Print ISSN 0001-706X
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 206
Article Number 105444
DOI https://doi.org/10.1016/j.actatropica.2020.105444
Keywords Leishmania; Drug screening; Bioluminescent reporters; Bioimaging; NanoLuc; Red-shifted luciferase
Publisher URL https://doi.org/10.1016/j.actatropica.2020.105444

Files







You might also like



Downloadable Citations