Artificial miRNAs targeting CAG repeat expansion in ORFs cause rapid deadenylation and translation inhibition of mutant transcripts

• DOI: 10.1007/s00018-020-03596-7
• Publication Date: 2020-07-21T20:02:46Z
• Authors: Adam Ciesiolka; Anna Stroynowska-...; Paweł Joachimiak; Agata Ciolak; Emilia Kozlowska; Michal Michalak; Magdalena Dabrowska; Marta Olejniczak; Katarzyna D. Racz...; Dominika Zielinska; Magdalena Wozna-W...; Wlodzimierz J. Kr...; Agnieszka Fiszer
• Keywords: ORFS

Abstract Polyglutamine (polyQ) diseases are incurable neurological disorders caused by CAG repeat expansion in the open reading frames (ORFs) of specific genes. This type mutation HTT gene is responsible for Huntington’s disease (HD). repeat-targeting artificial miRNAs (art-miRNAs) were shown as attractive therapeutic approach polyQ they allele-selective decrease level mutant proteins. Here, using models, we aimed to demonstrate how miRNA-based expression regulation dependent on target sequence features. We show that silencing efficiency and selectivity art-miRNAs influenced localization tract within transcript context. Furthermore, reveal events leading downregulation proteins found very rapid activation translational repression deadenylation. Slicer-activity AGO2 was dispensable this process, determined knockout cells generated with CRISPR-Cas9 technology. also showed highly huntingtin human HD neural progenitors (NPs). Taken together, art-miRNA activity may serve a model cooperative targeting ORF regions endogenous miRNAs.