2'-O-(N-(Aminoethyl)carbamoyl)methyl Modification Allows for Lower Phosphorothioate Content in Splice-Switching Oligonucleotides with Retained Activity.

Content Provider	Europe PMC
Author	Honcharenko, Dmytro Rocha, Cristina S.J. Lundin, Karin E. Maity, Jyotirmoy Milton, Stefan Tedebark, Ulf Murtola, Merita Honcharenko, Malgorzata Slaitas, Andis Smith, C.I. Edvard Zain, Rula Strömberg, Roger
Copyright Year	2022
Abstract	2′-O-(N-(Aminoethyl)carbamoyl)methyl (2′-O-AECM)-modified oligonucleotides (ONs) and their mixmers with 2′-O-methyl oligonucleotides (2′-OMe ONs) with phosphodiester linkers as well as with partial and full phosphorothioate (PS) inclusion were synthesized and functionally evaluated as splice-switching oligonucleotides in several different reporter cell lines originating from different tissues. This was enabled by first preparing the AECM-modified A, C, G and U, which required a different strategy for each building block. The AECM modification has previously been shown to provide high resistance to enzymatic degradation, even without PS linkages. It is therefore particularly interesting and unprecedented that the 2′-O-AECM ONs are shown to have efficient splice-switching activity even without inclusion of PS linkages and found to be as effective as 2′-OMe PS ONs. Importantly, the PS linkages can be partially included, without any significant reduction in splice-switching efficacy. This suggests that AECM modification has the potential to be used in balancing the PS content of ONs. Furthermore, conjugation of 2′-O-AECM ONs to an endosomal escape peptide significantly increased splice-switching suggesting that this effect could possibly be due to an increase in uptake of ON to the site of action.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC9221157&blobtype=pdf
Page Count	13
ISSN	21593337
Volume Number	32
DOI	10.1089/nat.2021.0086
PubMed Central reference number	PMC9221157
Issue Number	3
PubMed reference number	35238623
Journal	Nucleic Acid Therapeutics [Nucleic Acid Ther]
e-ISSN	21593345
Language	English
Publisher	Mary Ann Liebert, Inc., publishers
Publisher Date	2022-03-02
Publisher Place	USA
Access Restriction	Open
Rights License	This Open Access article is distributed under the terms of the Creative Commons License [CC-BY] (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © Dmytro Honcharenko et al., 2022; Published by Mary Ann Liebert, Inc.
Subject Keyword	2′-O-(N-(aminoethyl)carbamoyl)methyl modification splice-switching oligonucleotides cellular uptake cell-penetrating ONs phosphorothioate linkage oligonucleotide–peptide conjugate
Content Type	Text
Resource Type	Article
Subject	Genetics Drug Discovery Molecular Biology Biochemistry Molecular Medicine