An ultrasensitive NanoLuc-based luminescence system for monitoring Plasmodium berghei throughout its life cycle

De Niz Hidalgo, Mariana Isabel; Limenitakis Stanway, Rebecca; Wacker, Rahel Corina; Keller, Derya; Heussler, Volker T. (2016). An ultrasensitive NanoLuc-based luminescence system for monitoring Plasmodium berghei throughout its life cycle. Malaria journal, 15(1), p. 232. BioMed Central 10.1186/s12936-016-1291-9

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Bioluminescence imaging is widely used for cell-based assays and animal imaging studies, both in biomedical research and drug development. Its main advantages include its high-throughput applicability, affordability, high sensitivity, operational simplicity, and quantitative outputs. In malaria research, bioluminescence has been used for drug discovery in vivo and in vitro, exploring host-pathogen interactions, and studying multiple aspects of Plasmodium biology. While the number of fluorescent proteins available for imaging has undergone a great expansion over the last two decades, enabling simultaneous visualization of multiple molecular and cellular events, expansion of available luciferases has lagged behind. The most widely used bioluminescent probe in malaria research is the Photinus pyralis firefly luciferase, followed by the more recently introduced Click-beetle and Renilla luciferases. Ultra-sensitive imaging of Plasmodium at low parasite densities has not been previously achieved. With the purpose of overcoming these challenges, a Plasmodium berghei line expressing the novel ultra-bright luciferase enzyme NanoLuc, called PbNLuc has been generated, and is presented in this work.
NanoLuc shows at least 150 times brighter signal than firefly luciferase in vitro, allowing single parasite detection in mosquito, liver, and sexual and asexual blood stages. As a proof-of-concept, the PbNLuc parasites were used to image parasite development in the mosquito, liver and blood stages of infection, and to specifically explore parasite liver stage egress, and pre-patency period in vivo.
PbNLuc is a suitable parasite line for sensitive imaging of the entire Plasmodium life cycle. Its sensitivity makes it a promising line to be used as a reference for drug candidate testing, as well as the characterization of mutant parasites to explore the function of parasite proteins, host-parasite interactions, and the better understanding of Plasmodium biology. Since the substrate requirements of NanoLuc are different from those of firefly luciferase, dual bioluminescence imaging for the simultaneous characterization of two lines, or two separate biological processes, is possible, as demonstrated in this work.

Item Type:

Journal Article (Original Article)


08 Faculty of Science > Department of Biology > Institute of Cell Biology
08 Faculty of Science > Department of Biology > Institute of Cell Biology > Malaria

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

De Niz Hidalgo, Mariana Isabel, Limenitakis, Rebecca Rachel, Wacker, Rahel Corina, Keller, Derya, Heussler, Volker


500 Science > 570 Life sciences; biology




BioMed Central


[81] Evimalar ; [4] Swiss National Science Foundation




Volker Heussler

Date Deposited:

01 Jun 2016 14:18

Last Modified:

02 Mar 2023 23:27

Publisher DOI:


PubMed ID:





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