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New drugs are needed to overcome the toxicity, administration, low efficacy, and resistance issues of the current chemotherapy. Robust, simple, and economical high\u2010throughput, whole\u2010cell\u2010based assays are required to accelerate the identification of novel chemical entities. With this aim, we generated a bioluminescent cell line of the bloodstream stage of <jats:italic>Trypanosoma brucei brucei<\/jats:italic> and established a screening assay. Trypanosomes were stably transfected to constitutively express a thermostable red\u2010shifted luciferase. The growth phenotype and drug sensitivity of the reporter cell line were essentially identical to that of the parental cell line. The endogenous luciferase activity, measured by a simple bioluminescence assay, proved to be proportional to parasite number and metabolic status. The assay, optimized to detect highly potent compounds in a 96\u2010well\u2010plate format, was validated by screening a small compound library (inter\u2010assay values for Z' factor and coefficient variation were 0.77 and 5.8%, respectively). With a hit\u2010confirmation ratio of ~97%, the assay was potent enough to identify several hits with EC<jats:sub>50<\/jats:sub>\u2009\u2264\u200910\u00a0\u03bcM. Preliminary tests indicated that the assay can be scaled up to a 384\u2010well\u2010plate format without compromising its robustness. In summary, we have generated reporter trypanosomes and a simple, robust, and affordable bioluminescence screening assay with great potential to speed up the early\u2010phase drug discovery against African trypanosomes.<\/jats:p>","DOI":"10.1002\/ddr.21634","type":"journal-article","created":{"date-parts":[[2020,1,20]],"date-time":"2020-01-20T09:55:53Z","timestamp":1579514153000},"page":"253-263","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["A simple, robust, and affordable bioluminescent assay for drug discovery against infective African trypanosomes"],"prefix":"10.1002","volume":"83","author":[{"given":"Diego","family":"Ben\u00edtez","sequence":"first","affiliation":[{"name":"Group Redox Biology of Trypanosomes Institut Pasteur de Montevideo  Montevideo Uruguay"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Estefania","family":"Dibello","sequence":"additional","affiliation":[{"name":"Group Redox Biology of Trypanosomes Institut Pasteur de Montevideo  Montevideo Uruguay"},{"name":"Laboratorio de S\u00edntesis Org\u00e1nica, Departamento de Qu\u00edmica Org\u00e1nica, Facultad de Qu\u00edmica Universidad de la Rep\u00fablica  Montevideo Uruguay"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariana","family":"Bonilla","sequence":"additional","affiliation":[{"name":"Group Redox Biology of Trypanosomes Institut Pasteur de Montevideo  Montevideo Uruguay"},{"name":"Laboratorio de Fisicoqu\u00edmica Biol\u00f3gica y Laboratorio de Enzimolog\u00eda, Instituto de Qu\u00edmica Biol\u00f3gica, Facultad de Ciencias Universidad de la Rep\u00fablica  Montevideo Uruguay"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5000-1333","authenticated-orcid":false,"given":"Marcelo A.","family":"Comini","sequence":"additional","affiliation":[{"name":"Group Redox Biology of Trypanosomes Institut Pasteur de Montevideo  Montevideo Uruguay"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,1,20]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molbiopara.2004.10.002"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijpddr.2012.02.002"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2009.09.009"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dku393"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.freeradbiomed.2004.02.008"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1017\/S003118201300142X"},{"key":"e_1_2_7_8_1","first-page":"270","article-title":"Der Nachweis der hydrosulfite (sulfoxylate) und des nachierenden Wasserstoffs mit Resazurin. 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