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Cellular uptake and trafficking of antisense oligonucleotides

Abstract

Antisense oligonucleotides (ASOs) modified with phosphorothioate (PS) linkages and different 2′ modifications can be used either as drugs (e.g., to treat homozygous familial hypercholesterolemia and spinal muscular atrophy) or as research tools to alter gene expression. PS-ASOs can enter cells without additional modification or formulation and can be designed to mediate sequence-specific cleavage of different types of RNA (including mRNA and non-coding RNA) targeted by endogenous RNase H1. Although PS-ASOs function in both the cytoplasm and nucleus, localization to different subcellular regions can affect their therapeutic potency. Cellular uptake and intracellular distribution of PS ASOs are mediated by protein interactions. The main proteins involved in these processes have been identified, and intracellular sites in which PS ASOs are active, or inactive, cataloged.

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Figure 1: Pathways of PS-ASO uptake and intracellular trafficking.
Figure 2: PS-ASOs interact with intracellular proteins.
Figure 3: PS-ASO localization in nuclear and cytoplasmic foci.
Figure 4: Kinetics of PS-ASO localization and antisense activity.

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Acknowledgements

This work is supported by internal funding from Ionis Pharmaceuticals. The authors wish to thank R. Crooke for helpful comments, T. Kniss and D. Parrett for editing and administrative assistance, and T. Reigle for help in figure preparation.

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Correspondence to Stanley T Crooke.

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This work was funded by Ionis Pharmaceuticals. All authors are employees of Ionis Pharmaceuticals.

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Crooke, S., Wang, S., Vickers, T. et al. Cellular uptake and trafficking of antisense oligonucleotides. Nat Biotechnol 35, 230–237 (2017). https://doi.org/10.1038/nbt.3779

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