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Fine Tuning of Phos...
Fine Tuning of Phosphorothioate Inclusion in 2'-O-Methyl Oligonucleotides Contributes to Specific Cell Targeting for Splice-Switching Modulation
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Aoki, Y (författare)
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Rocha, CSJ (författare)
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- Lehto, T (författare)
- Karolinska Institutet
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Miyatake, S (författare)
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- Johansson, H (författare)
- Karolinska Institutet
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Hashimoto, Y (författare)
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- Nordin, JZ (författare)
- Karolinska Institutet
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Mager, I (författare)
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Aoki, M (författare)
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Graham, M (författare)
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Sathyaprakash, C (författare)
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Roberts, TC (författare)
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Wood, MJA (författare)
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Behlke, MA (författare)
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- El Andaloussi, S (författare)
- Karolinska Institutet
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(creator_code:org_t)
- 2021-10-13
- 2021
- Engelska.
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Ingår i: Frontiers in physiology. - : Frontiers Media SA. - 1664-042X. ; 12, s. 689179-
- Relaterad länk:
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https://doi.org/10.3...
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http://kipublication...
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https://doi.org/10.3...
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Abstract
Ämnesord
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- 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.
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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- Av författaren/redakt...
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Aoki, Y
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Rocha, CSJ
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Lehto, T
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Miyatake, S
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Johansson, H
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Hashimoto, Y
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visa fler...
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Nordin, JZ
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Mager, I
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Aoki, M
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Graham, M
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Sathyaprakash, C
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Roberts, TC
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Wood, MJA
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Behlke, MA
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El Andaloussi, S
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visa färre...
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Frontiers in phy ...
- Av lärosätet
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Karolinska Institutet