Whole worm microRNA silencing in cestodes

GRECCO, ANDRES; MACCHIAROLI, NATALIA; CUCHER, MARCELA; ROSENZVIT, MARA

Congreso; XXXII Molecular Parasitology Meeting; 2021

Whole worm microRNA silencing in cestodesAutores: Andrés Grecco, Natalia Macchiaroli, Marcela Cucher and Mara RosenzvitInstitución: Laboratorio Biología Molecular de Hidatidosis, Instituto de Microbiología y Parasitología Médica (IMPaM), Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Facultad de Medicina, Universidad de Buenos Aires (UBA), Buenos Aires, Argentina. Contacto: andresgrecco1@gmail.comMesocestoides vogae (syn.M.corti) is a platyhelminth parasite validated as a laboratory model of the Cestoda class. Cestodes include species such as Echinococcus granulosus that cause Tropical Neglected Diseases (NTDs). Although genomic information is available for several of these parasites, functional postgenomic strategies are very limited so far. microRNAs (miRNAs) are small non-coding RNAs with a main role in regulation of gene expression. miR-71 is highly expressed in cestodes, is absent in vertebrates and targets essential parasite genes. Thus, miR-71 represents a potential selective drug target to treat cestodiases. Here, we aim to knockdown miR-71 in whole larvae (tetrathyridia) from M. vogae. To this end, a strategy based on antimiRs was selected. AntimiRs are single-stranded oligonucleotides complementary to the miRNA, which block miRNA function. First, transfections with increasing concentrations of a fluorescent scramble oligonucleotide were performed on cultured tetrathyridia and analyzed through confocal microscopy to determine the best uptake conditions. Administration of antimiR-71 at 3 and 9 µM resulted in miR-71 knockdown of 68% and 83% respectively, measured by RT-qPCR. Analysis of miRNA silencing effects on parasite development is underway. This result provides a strategy to study miRNA function in cestodes and to analyze miRNA potential as drug targets against NTDs.