{"entity": "publication", "iuid": "3c57a24495f04ec3b098f70505ca3426", "timestamp": "2026-08-17T17:24:50.300Z", "links": {"self": {"href": "https://publications.scilifelab.se/publication/3c57a24495f04ec3b098f70505ca3426.json"}, "display": {"href": "https://publications.scilifelab.se/publication/3c57a24495f04ec3b098f70505ca3426"}}, "title": "Fine Tuning of Phosphorothioate Inclusion in 2'-O-Methyl Oligonucleotides Contributes to Specific Cell Targeting for Splice-Switching Modulation.", "authors": [{"family": "Aoki", "given": "Yoshitsugu", "initials": "Y"}, {"family": "Rocha", "given": "Cristina S J", "initials": "CSJ"}, {"family": "Lehto", "given": "Taavi", "initials": "T"}, {"family": "Miyatake", "given": "Shouta", "initials": "S"}, {"family": "Johansson", "given": "Henrik", "initials": "H"}, {"family": "Hashimoto", "given": "Yasumasa", "initials": "Y"}, {"family": "Nordin", "given": "Joel Z", "initials": "JZ"}, {"family": "Mager", "given": "Imre", "initials": "I"}, {"family": "Aoki", "given": "Misako", "initials": "M"}, {"family": "Graham", "given": "McClorey", "initials": "M"}, {"family": "Sathyaprakash", "given": "Chaitra", "initials": "C"}, {"family": "Roberts", "given": "Thomas C", "initials": "TC"}, {"family": "Wood", "given": "Matthew J A", "initials": "MJA"}, {"family": "Behlke", "given": "Mark A", "initials": "MA"}, {"family": "Andaloussi", "given": "Samir El", "initials": "SE"}], "type": "journal article", "published": "2021-10-13", "journal": {"title": "Front Physiol", "issn": "1664-042X", "volume": "12", "pages": "689179", "issn-l": null}, "abstract": "Splice-switching antisense oligonucleotide- (SSO-) mediated correction of framedisrupting mutation-containing premessenger RNA (mRNA) transcripts using exon skipping is a highly promising treatment method for muscular diseases such as Duchenne muscular dystrophy (DMD). Phosphorothioate (PS) chemistry, a commonly used oligonucleotide modification, has been shown to increase the stability of and improve the pharmacokinetics of SSOs. However, the effect of PS inclusion in 2'-O-methyl SSOs (2OMe) on cellular uptake and splice switching is less well-understood. At present, we demonstrate that the modification of PS facilitates the uptake of 2OMe in H2k-mdx myoblasts. Furthermore, we found a dependency of SSO nuclear accumulation and high splice-switching activity on PS inclusion in 2OMe (2OMePS), as tested in various reporter cell lines carrying pLuc/705. Increased exon-inclusion activity was observed in muscle, neuronal, liver, and bone cell lineages via both the gymnotic uptake and lipofection of 2OMePS. Using the photoactivatable ribonucleoside-enhanced crosslinking and a subsequent proteomic approach, we identified several 2OMePS-binding proteins, which are likely to play a role in the trafficking of 2OMePS to the nucleus. Ablation of one of them, Ncl by small-interfering RNA (siRNA) enhanced 2OMePS uptake in C2C12 myoblasts and upregulated luciferase RNA splicing in the HeLa Luc/705 reporter cell line. Overall, we demonstrate that PS inclusion increases nuclear delivery and splice switching in muscle, neuronal, liver, and bone cell lineages and that the modulation of 2OMePS-binding partners may improve SSO delivery.", "doi": "10.3389/fphys.2021.689179", "pmid": "34721051", "labels": {"Global Proteomics and Proteogenomics": "Service"}, "xrefs": [{"db": "pmc", "key": "PMC8548633"}], "notes": [], "created": "2022-03-29T10:44:48.421Z", "modified": "2022-03-29T10:44:48.450Z"}