Generation and Characterization of a Dual-Reporter Transgenic Leishmania braziliensis Line Expressing eGFP and Luciferase

Rohit Sharma; Paulo S Silveira-Mattos; Vinicius C Ferreira; Francys A Rangel; Laíse B Oliveira; Fabiana S Celes; Sayonara M Viana; Mary E Wilson; Camila I de Oliveira

doi:10.3389/fcimb.2019.00468

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Generation and Characterization of a Dual-Reporter Transgenic Leishmania braziliensis Line Expressing eGFP and Luciferase

Journal article

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Peer reviewed

Generation and Characterization of a Dual-Reporter Transgenic Leishmania braziliensis Line Expressing eGFP and Luciferase

Rohit Sharma, Paulo S Silveira-Mattos, Vinicius C Ferreira, Francys A Rangel, Laíse B Oliveira, Fabiana S Celes, Sayonara M Viana, Mary E Wilson and Camila I de Oliveira

Frontiers in cellular and infection microbiology, Vol.9, pp.468-468

2019

DOI: 10.3389/fcimb.2019.00468

PMCID: PMC6987073

PMID: 32039047

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Abstract

In this study, we generated a transgenic strain of , an etiological agent associated with a diversity of clinical manifestations of leishmaniasis ranging from localized cutaneous to mucocutaneous to disseminated disease. Transgenic parasites expressing reporter proteins are valuable tools for studies of parasite biology, host-pathogen interactions, and anti-parasitic drug development. To this end, we constructed an line stably expressing the reporters eGFP and luciferase (eGFP-LUC . The integration cassette co-expressing the two reporters was targeted to the ribosomal locus ( ) of the parasite genome. Transgenic parasites were characterized for their infectivity and stability both and . Parasite maintenance in axenic long-term culture in the absence of selective drugs did not alter expression of the two reporters or infection of BALB/c mice, indicating stability of the integrated cassette. Infectivity of eGFP-LUC, , both and was similar to that obtained with the parental wild type strain. The possibility of tracking and quantification using fluorescence and luminescence broadens the scope of research involving this neglected species, despite its importance in terms of public health concerning the leishmaniasis burden.

Animals

Disease Models, Animal

Genes, Reporter

Genomic Instability

Green Fluorescent Proteins - analysis

Green Fluorescent Proteins - genetics

Leishmania braziliensis - genetics

Leishmania braziliensis - metabolism

Leishmaniasis, Cutaneous - parasitology

Luciferases - analysis

Luciferases - genetics

Luminescent Agents - analysis

Mice, Inbred BALB C

Recombinant Proteins - analysis

Recombinant Proteins - genetics

Staining and Labeling - methods

Details

Title: Subtitle: Generation and Characterization of a Dual-Reporter Transgenic Leishmania braziliensis Line Expressing eGFP and Luciferase
Creators: Rohit Sharma - Fundação Oswaldo Cruz
Paulo S Silveira-Mattos - Fundação Oswaldo Cruz
Vinicius C Ferreira - Fundação Oswaldo Cruz
Francys A Rangel - Fundação Oswaldo Cruz
Laíse B Oliveira - Fundação Oswaldo Cruz
Fabiana S Celes - Fundação Oswaldo Cruz
Sayonara M Viana - Fundação Oswaldo Cruz
Mary E Wilson - University of Iowa
Camila I de Oliveira - Fundação Oswaldo Cruz
Resource Type: Journal article
Publication Details: Frontiers in cellular and infection microbiology, Vol.9, pp.468-468
DOI: 10.3389/fcimb.2019.00468
PMID: 32039047
PMCID: PMC6987073
NLM abbreviation: Front Cell Infect Microbiol
ISSN: 2235-2988
eISSN: 2235-2988
Grant note: R01 AI045540 / NIAID NIH HHS R01 AI076233 / NIAID NIH HHS I01 BX000536 / BLRD VA I01 BX001983 / BLRD VA
Language: English
Date published: 2019
Academic Unit: Microbiology and Immunology; Infectious Diseases; International Programs; Epidemiology; Internal Medicine
Record Identifier: 9984297434702771

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