Artificial miRNAs targeting CAG repeat expansion in ORFs cause rapid deadenylation and translation inhibition of mutant transcripts
| Autor: | Wlodzimierz J. Krzyzosiak, Agata Ciolak, Michał Michalak, Agnieszka Fiszer, Marta Olejniczak, Katarzyna Dorota Raczynska, Dominika Zielinska, Magdalena Wozna-Wysocka, Adam Ciesiolka, Emilia Kozlowska, Magdalena Dabrowska, Paweł Joachimiak, Anna Stroynowska-Czerwinska |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Huntingtin
Mutant CAG repeats Biology medicine.disease_cause 03 medical and health sciences Cellular and Molecular Neuroscience Polyglutamine diseases Gene Knockout Techniques Open Reading Frames 0302 clinical medicine medicine Gene silencing Humans ORFS Molecular Biology Translational inhibition Alleles 030304 developmental biology miRNA Pharmacology Regulation of gene expression 0303 health sciences Mutation Huntingtin Protein Cell Biology Cell biology Open reading frame MicroRNAs Huntington Disease Protein Biosynthesis Argonaute Proteins Molecular Medicine Original Article RNA Interference CRISPR-Cas Systems Trinucleotide repeat expansion Peptides Trinucleotide Repeat Expansion 030217 neurology & neurosurgery Huntington’s disease |
| Zdroj: | Cellular and Molecular Life Sciences |
| ISSN: | 1420-9071 1420-682X |
| Popis: | Polyglutamine (polyQ) diseases are incurable neurological disorders caused by CAG repeat expansion in the open reading frames (ORFs) of specific genes. This type of mutation in the HTT gene is responsible for Huntington’s disease (HD). CAG repeat-targeting artificial miRNAs (art-miRNAs) were shown as attractive therapeutic approach for polyQ disorders as they caused allele-selective decrease in the level of mutant proteins. Here, using polyQ disease models, we aimed to demonstrate how miRNA-based gene expression regulation is dependent on target sequence features. We show that the silencing efficiency and selectivity of art-miRNAs is influenced by the localization of the CAG repeat tract within transcript and the specific sequence context. Furthermore, we aimed to reveal the events leading to downregulation of mutant polyQ proteins and found very rapid activation of translational repression and HTT transcript deadenylation. Slicer-activity of AGO2 was dispensable in this process, as determined in AGO2 knockout cells generated with CRISPR-Cas9 technology. We also showed highly allele-selective downregulation of huntingtin in human HD neural progenitors (NPs). Taken together, art-miRNA activity may serve as a model of the cooperative activity and targeting of ORF regions by endogenous miRNAs. Electronic supplementary material The online version of this article (10.1007/s00018-020-03596-7) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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