title: A non-commercial approach for the generation of transgenic Leishmania tarentolae and its application in antileishmanial drug discovery.
abstract: Leishmaniasis is a parasitic infection caused by several species of the genus Leishmania that is considered as a neglected disease. Drug development process requires a robust and updated high-throughput technology to the evaluation of candidate compounds that imply the manipulation of the pathogenic species of the parasite in the laboratory. Therefore, it is restricted to trained personal and level II biosafety environments. However, it has been established the utility of Leishmania tarentolae as a model for in vitro screening of antileishmanial agents without the necessity of level II biosafety setups. In parallel the transfection of Leishmania parasites with reporter genes as the eGFP using non-commercial integration vectors like the pIRmcs3(-) has proved to be a powerful tool for the implementation of semi automatized high-throughput platforms for the evaluation of antileishmanial compounds. Here we report the generation of a new L. tarentolae strain overexpressing the eGFP gene harboured by the non-commercial vector pIR3(-). We also demonstrate its utility for the semi-automatized screening of antileshmanial compounds in intracellular forms of the L. tarentolae parasite.
authors.affiliation: PECET,Medical Research Institute,School of Medicine,University of Antioquia,Medellín,Colombia.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Pineda
authors.affiliation: PECET,Medical Research Institute,School of Medicine,University of Antioquia,Medellín,Colombia.
authors.forename: Yesenia
authors.initials: Y
authors.lastname: Valencia
authors.affiliation: PECET,Medical Research Institute,School of Medicine,University of Antioquia,Medellín,Colombia.
authors.forename: María F
authors.initials: MF
authors.lastname: Flórez
authors.affiliation: PECET,Medical Research Institute,School of Medicine,University of Antioquia,Medellín,Colombia.
authors.forename: Sergio A
authors.initials: SA
authors.lastname: Pulido
authors.affiliation: PECET,Medical Research Institute,School of Medicine,University of Antioquia,Medellín,Colombia.
authors.forename: Iván D
authors.initials: ID
authors.lastname: Vélez
authors.affiliation: PECET,Medical Research Institute,School of Medicine,University of Antioquia,Medellín,Colombia.
authors.forename: Sara M
authors.initials: SM
authors.lastname: Robledo
mesh_terms: D000981:Antiprotozoal Agents / Q000494:pharmacology* / Q000627:therapeutic use; D019295:Computational Biology; D055808:Drug Discovery; D005434:Flow Cytometry; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D057166:High-Throughput Screening Assays; D006801:Humans; D020128:Inhibitory Concentration 50; D007891:Leishmania / Q000187:drug effects / Q000235:genetics*; D007896:Leishmaniasis / Q000188:drug therapy / Q000469:parasitology*; D058069:Neglected Diseases / Q000188:drug therapy / Q000469:parasitology*; D030781:Organisms, Genetically Modified / Q000235:genetics*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012449:Safety; D020298:U937 Cells
chemical_list: D000981:Antiprotozoal Agents; D012337:RNA, Ribosomal, 18S; D049452:Green Fluorescent Proteins
keywords: Leishmania tarentolae; green fluorescent protein; pIRmsc3(−); transgenic parasites