期刊
NUCLEIC ACID THERAPEUTICS
卷 32, 期 3, 页码 221-233出版社
MARY ANN LIEBERT, INC
DOI: 10.1089/nat.2021.0086
关键词
2 '-O-(N-(aminoethyl)carbamoyl)methyl modification; splice-switching oligonucleotides; cellular uptake; cell-penetrating ONs; phosphorothioate linkage; oligonucleotide-peptide conjugate
资金
- Swedish Research Council [2012-5715, 2016-06155, 2016-03283, K2015-68X-11247-21-3, 2017-02131]
- Duchenne Parent Project Netherlands (DPP NL)
- Stockholm County Council (Stockholms lans landsting)
- Knut and Alice Wallenberg Foundation
- Swedish Research Council
- Center for Innovative Medicine
This study synthesized 2'-O-AECM-modified ONs and found that these modified ONs have efficient splice-switching activity even without inclusion of PS linkages. Furthermore, partial inclusion of PS linkages does not significantly reduce the splice-switching efficacy. In addition, conjugation of 2'-O-AECM ONs with an endosomal escape peptide increases splice-switching activity.
2 & PRIME;-O-(N-(Aminoethyl)carbamoyl)methyl (2 & PRIME;-O-AECM)-modified oligonucleotides (ONs) and their mixmers with 2 & PRIME;-O-methyl oligonucleotides (2 & PRIME;-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 & PRIME;-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 & PRIME;-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 & PRIME;-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.
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