Dhorne-Pollet, SophieFitzpatrick, ChristopherDa Costa, BrunoBourgon, ClaraEléouët, Jean-FrançoisMeunier, NicolasBurzio, Verónica A.Delmas, BernardBarrey, Eric2023-04-172023-04-172022-10Frontiers in Microbiology Volume 1326 October 2022 Article number 9152021664-302Xhttps://repositorio.unab.cl/xmlui/handle/ria/48639Indexación: ScopusThe ongoing COVID-19 pandemic continues to pose a need for new and efficient therapeutic strategies. We explored antisense therapy using oligonucleotides targeting the severe acute respiratory syndrome coronavirus (SARS-CoV-2) genome. We predicted in silico four antisense oligonucleotides (ASO gapmers with 100% PTO linkages and LNA modifications at their 5′ and 3′ends) targeting viral regions ORF1a, ORF1b, N and the 5′UTR of the SARS-CoV-2 genome. Efficiency of ASOs was tested by transfection in human ACE2-expressing HEK-293T cells and monkey VeroE6/TMPRSS2 cells infected with SARS-CoV-2. The ORF1b-targeting ASO was the most efficient, with a 71% reduction in the number of viral genome copies. N- and 5′UTR-targeting ASOs also significantly reduced viral replication by 55 and 63%, respectively, compared to non-related control ASO (ASO-C). Viral titration revealed a significant decrease in SARS-CoV-2 multiplication both in culture media and in cells. These results show that anti-ORF1b ASO can specifically reduce SARS-CoV-2 genome replication in vitro in two different cell infection models. The present study presents proof-of concept of antisense oligonucleotide technology as a promising therapeutic strategy for COVID-19. Copyright © 2022 Dhorne-Pollet, Fitzpatrick, Da Costa, Bourgon, Eléouët, Meunier, Burzio, Delmas and Barrey.enASOCoronavirus; oligonucleotide antisense therapyRNA therapyRNA virusSARS-CoV-2SARS-CoV-2Antisense therapy RNA virusArtículoAtribución 4.0 Internacional (CC BY 4.0)10.3389/fmicb.2022.915202