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Therefore, new drugs and assays to screen and detect novel active compounds against leishmaniasis are urgently needed. We thus validated\n                    <jats:italic>Leishmania braziliensis<\/jats:italic>\n                    (Lb) and\n                    <jats:italic>Leishmania infantum<\/jats:italic>\n                    (Li) that constitutively express the tandem tomato red fluorescent protein (\n                    <jats:italic>tdTomato<\/jats:italic>\n                    ) as a model for large-scale screens of anti-\n                    <jats:italic>Leishmania<\/jats:italic>\n                    compounds. Confocal microscopy of\n                    <jats:italic>Lb<\/jats:italic>\n                    and\n                    <jats:italic>Li::tdTomato<\/jats:italic>\n                    revealed red fluorescence distributed throughout the entire parasite, including the flagellum, and flow cytometry confirmed that the parasites emitted intense fluorescence. We evaluated the infectivity of cloned promastigotes and amastigotes constitutively expressing\n                    <jats:italic>tdTomato<\/jats:italic>\n                    , their growth profiles in THP-1 macrophages, and susceptibility to trivalent antimony, amphotericin, and miltefosine\n                    <jats:italic>in vitro<\/jats:italic>\n                    . The phenotypes of mutant and wild-type parasites were similar, indicating that the constitutive expression of\n                    <jats:italic>tdTomato<\/jats:italic>\n                    did not interfere with the evaluated parameters. We applied our validated model to a repositioning strategy and assessed the susceptibility of the parasites to eight commercially available drugs. We also screened 32 natural plant and fungal extracts and 10 pure substances to reveal new active compounds. The infectivity and Glucantime treatment efficacy of BALB\/c mice and golden hamsters infected with\n                    <jats:italic>Lb<\/jats:italic>\n                    and\n                    <jats:italic>Li::tdTomato<\/jats:italic>\n                    mutant lines, respectively, were very similar compared to animals infected with wild-type parasites. Standardizing our methodology would offer more rapid, less expensive, and easier assays to screen of compounds against\n                    <jats:italic>L. braziliensis<\/jats:italic>\n                    and\n                    <jats:italic>L. infantum in vitro<\/jats:italic>\n                    and\n                    <jats:italic>in vivo<\/jats:italic>\n                    . Our method could also enhance the discovery of active compounds for treating leishmaniasis.\n                  <\/jats:p>","DOI":"10.1128\/aac.00509-23","type":"journal-article","created":{"date-parts":[[2023,12,8]],"date-time":"2023-12-08T09:00:20Z","timestamp":1702026020000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":14,"title":["Anti-\n                    <i>Leishmania<\/i>\n                    compounds can be screened using\n                    <i>Leishmania<\/i>\n                    spp. expressing red fluorescence (\n                    <i>tdTomato<\/i>\n                    )"],"prefix":"10.1128","volume":"68","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0828-790X","authenticated-orcid":false,"given":"Mariza Gabriela Faleiro de Moura Lodi","family":"Cruz","sequence":"first","affiliation":[{"name":"Gen\u00f4mica Funcional de Parasitos, Instituto Ren\u00e9 Rachou, Funda\u00e7\u00e3o Oswaldo Cruz FIOCRUZ Minas , Belo Horizonte, Minas Gerais, Brazil"},{"name":"Qu\u00edmica de Produtos Naturais Bioativos, Instituto Ren\u00e9 Rachou, Funda\u00e7\u00e3o Oswaldo Cruz FIOCRUZ Minas , Belo Horizonte, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9791-6554","authenticated-orcid":false,"given":"Ana Maria Murta","family":"Santi","sequence":"additional","affiliation":[{"name":"Gen\u00f4mica Funcional de Parasitos, Instituto Ren\u00e9 Rachou, Funda\u00e7\u00e3o Oswaldo Cruz FIOCRUZ Minas , Belo Horizonte, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eliane de","family":"Morais-Teixeira","sequence":"additional","affiliation":[{"name":"Pesquisa Cl\u00ednica e Pol\u00edticas P\u00fablicas em Doen\u00e7as Infecto-Parasit\u00e1rias, Instituto Ren\u00e9 Rachou, Funda\u00e7\u00e3o Oswaldo Cruz FIOCRUZ Minas , Belo Horizonte, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alisson Samuel Portes","family":"Caldeira","sequence":"additional","affiliation":[{"name":"Qu\u00edmica de Produtos Naturais Bioativos, Instituto Ren\u00e9 Rachou, Funda\u00e7\u00e3o Oswaldo Cruz FIOCRUZ Minas , Belo Horizonte, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ezequias Pessoa de","family":"Siqueira","sequence":"additional","affiliation":[{"name":"Qu\u00edmica de Produtos Naturais Bioativos, Instituto Ren\u00e9 Rachou, Funda\u00e7\u00e3o Oswaldo Cruz FIOCRUZ Minas , Belo Horizonte, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0042-5178","authenticated-orcid":false,"given":"Edward","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Gen\u00f4mica Funcional de Parasitos, Instituto Ren\u00e9 Rachou, Funda\u00e7\u00e3o Oswaldo Cruz FIOCRUZ Minas , Belo Horizonte, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0582-3968","authenticated-orcid":false,"given":"T\u00e2nia Maria de Almeida","family":"Alves","sequence":"additional","affiliation":[{"name":"Qu\u00edmica de Produtos Naturais Bioativos, Instituto Ren\u00e9 Rachou, Funda\u00e7\u00e3o Oswaldo Cruz FIOCRUZ Minas , Belo Horizonte, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8523-2155","authenticated-orcid":true,"given":"Silvane Maria Fonseca","family":"Murta","sequence":"additional","affiliation":[{"name":"Gen\u00f4mica Funcional de Parasitos, Instituto Ren\u00e9 Rachou, Funda\u00e7\u00e3o Oswaldo Cruz FIOCRUZ Minas , Belo Horizonte, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","volume-title":"Leishmaniasis","author":"World health organization","year":"2023","unstructured":"World health organization . 2023. 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