Author | Sharma, Rohit | |
Author | Mattos, Paulo Sérgio de Morais da Silveira | |
Author | Ferreira, Vinicius C. | |
Author | Rangel, Francys A. | |
Author | Oliveira, Laíse B. | |
Author | Celes, Fabiana S. | |
Author | Viana, Sayonara M. | |
Author | Wilson, Mary E. | |
Author | Oliveira, Camila Indiani de | |
Access date | 2020-03-10T11:48:34Z | |
Available date | 2020-03-10T11:48:34Z | |
Document date | 2019 | |
Citation | SHARMA, Rohit et al. Generation and Characterization of a Dual-Reporter Transgenic Leishmania braziliensis Line Expressing eGFP and Luciferase. Frontiers in Cellular and Infection Microbiology, v. 9, p. 1-10, 2020. | pt_BR |
ISSN | 2235-2988 | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/40265 | |
Sponsorship | Coordenação
de Aperfeiçoamento de pessoal de Nível Superior—Brasil
(CAPES)—Finance Code 001. LO was financed by a CNPq
fellowship (Iniciação Científica). CO was a senior investigator
from CNPq. This work was funded in part by FIOCRUZ, and by
US National Institutes of Health grants R01 AI045540 and R01
AI076233 (MW), and by Department of Veterans’ Affairs Merit
Review Grants BX001983, BX000536 (MW). | pt_BR |
Language | eng | pt_BR |
Publisher | Frontiers Media | pt_BR |
Rights | open access | pt_BR |
Subject in Portuguese | L. braziliensis | pt_BR |
Subject in Portuguese | Transgênico | pt_BR |
Subject in Portuguese | Repórteres duplos | pt_BR |
Subject in Portuguese | Luciferase | pt_BR |
Subject in Portuguese | GFP | pt_BR |
Title | Generation and Characterization of a Dual-Reporter Transgenic Leishmania braziliensis Line Expressing eGFP and Luciferase | pt_BR |
Type | Article | pt_BR |
DOI | 10.3389/fcimb.2019.00468 | |
Abstract | In this study, we generated a transgenic strain of Leishmania braziliensis, 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 L. braziliensis line stably expressing the reporters eGFP and luciferase (eGFP-LUC L. braziliensis). The integration cassette co-expressing the two reporters was targeted to the ribosomal locus (SSU) of the parasite genome. Transgenic parasites were characterized for their infectivity and stability both in vitro and in vivo. 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, L. braziliensis, both in vivo and in vitro was similar to that obtained with the parental wild type strain. The possibility of L. braziliensis 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. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. | pt_BR |
Affilliation | University of Iowa, and the Veterans’ Affairs Medical Center. Departments of Microbiology and Immunology and Internal Medicine. Iowa City, IA, United States. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Instituto de Investigação em Imunologia. São Paulo, SP, Brasil. | pt_BR |
Subject | L. braziliensis | pt_BR |
Subject | Transgenic | pt_BR |
Subject | Dual reporters | pt_BR |
Subject | Luciferase | pt_BR |
Subject | GFP | pt_BR |