Evaluation of anhydrohexitol nucleic acid, cyclohexenyl nucleic acid andd-altritol nucleic acid-modified 2′-O-methyl RNA mixmer antisense oligonucleotides for exon skipping in vitro

2016 ◽  
Vol 52 (92) ◽  
pp. 13467-13470 ◽  
Author(s):  
Bao T. Le ◽  
Suxiang Chen ◽  
Mikhail Abramov ◽  
Piet Herdewijn ◽  
Rakesh N. Veedu
Keyword(s):  
Nucleic Acid ◽  
Antisense Oligonucleotides ◽  
Exon Skipping ◽  
Antisense Oligos

We have investigated the potential of anhydrohexitol, cyclohexenyl and altritol nucleic acid-modified antisense oligos in exon skipping, and found that they efficiently inducedDmdexon 23 skipping.

scholarly journals Rational Design of Short Locked Nucleic Acid-Modified 2′- O -Methyl Antisense Oligonucleotides for Efficient Exon-Skipping In Vitro

2017 ◽  
Vol 9 ◽  
pp. 155-161 ◽  
Author(s):  
Bao T. Le ◽  
Abbie M. Adams ◽  
Susan Fletcher ◽  
Stephen D. Wilton ◽  
Rakesh N. Veedu
Keyword(s):  
Nucleic Acid ◽  
Antisense Oligonucleotides ◽  
Rational Design ◽  
Exon Skipping ◽  
Locked Nucleic Acid

Investigation of twisted intercalating nucleic acid (TINA)-modified antisense oligonucleotides for splice modulation by induced exon-skipping in vitro

RSC Advances ◽  
2016 ◽  
Vol 6 (97) ◽  
pp. 95169-95172 ◽  
Author(s):  
Bao T. Le ◽  
Vyacheslav V. Filichev ◽  
Rakesh N. Veedu
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Antisense Oligonucleotides ◽  
Exon Skipping

We have investigated the applicability of twisted intercalating nucleic acids (TINA)-modified antisense oligonucleotides (AOs) in exon skipping. We found that TINA-modified AOs induced exon skipping.

scholarly journals Alpha-l-Locked Nucleic Acid-Modified Antisense Oligonucleotides Induce Efficient Splice Modulation In Vitro

2020 ◽  
Vol 21 (7) ◽  
pp. 2434
Author(s):  
Prithi Raguraman ◽  
Tao Wang ◽  
Lixia Ma ◽  
Per Trolle Jørgensen ◽  
Jesper Wengel ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Antisense Oligonucleotides ◽  
Exon Skipping ◽  
Initial Study ◽  
Locked Nucleic Acid ◽  
Mdx Mouse ◽  
Complementary Oligonucleotide ◽  
Low Cytotoxicity ◽  
Target Binding

Alpha-l-Locked nucleic acid (α-l-LNA) is a stereoisomeric analogue of locked nucleic acid (LNA), which possesses excellent biophysical properties and also exhibits high target binding affinity to complementary oligonucleotide sequences and resistance to nuclease degradations. Therefore, α-l-LNA nucleotides could be utilised to develop stable antisense oligonucleotides (AO), which can be truncated without compromising the integrity and efficacy of the AO. In this study, we explored the potential of α-l-LNA nucleotides-modified antisense oligonucleotides to modulate splicing by inducing Dmd exon-23 skipping in mdx mouse myoblasts in vitro. For this purpose, we have synthesised and systematically evaluated the efficacy of α-l-LNA-modified 2′-O-methyl phosphorothioate (2′-OMePS) AOs of three different sizes including 20mer, 18mer and 16mer AOs in parallel to fully-modified 2′-OMePS control AOs. Our results demonstrated that the 18mer and 16mer truncated AO variants showed slightly better exon-skipping efficacy when compared with the fully-23 modified 2′-OMePS control AOs, in addition to showing low cytotoxicity. As there was no previous report on using α-l-LNA-modified AOs in splice modulation, we firmly believe that this initial study could be beneficial to further explore and expand the scope of α-l-LNA-modified AO therapeutic molecules.

In vitro evaluation of hexitol nucleic acid antisense oligonucleotides

1999 ◽  
Author(s):  
Arthur Van Aerschot ◽  
Mark Vandermeeren ◽  
Johan Geysen ◽  
Walter Luyten ◽  
Marc Miller ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Antisense Oligonucleotides ◽  
In Vitro Evaluation

scholarly journals Nucleobase-modified antisense oligonucleotides containing 5-(phenyltriazol)-2′-deoxyuridine nucleotides induce exon-skipping in vitro

RSC Advances ◽  
2017 ◽  
Vol 7 (86) ◽  
pp. 54542-54545 ◽  
Author(s):  
Bao T. Le ◽  
Mick Hornum ◽  
Pawan K. Sharma ◽  
Poul Nielsen ◽  
Rakesh N. Veedu
Keyword(s):  
Antisense Oligonucleotides ◽  
Exon Skipping

We investigated the potential of nucleobase-modified antisense oligonucleotides to induce exon-skipping, and found that 5-(phenyltriazol)-2′-deoxyuridine-modified antisense oligonucleotides induced efficient exon-skipping in vitro.

scholarly journals Proof-of-Concept: Antisense Oligonucleotide Mediated Skipping of Fibrillin-1 Exon 52

2021 ◽  
Vol 22 (7) ◽  
pp. 3479
Author(s):  
Jessica M. Cale ◽  
Kane Greer ◽  
Sue Fletcher ◽  
Steve D. Wilton
Keyword(s):  
Antisense Oligonucleotide ◽  
Antisense Oligonucleotides ◽  
Exon Skipping ◽  
Mrna Splicing ◽  
Immunofluorescent Staining ◽  
Proof Of Concept ◽  
Connective Tissue Disorders ◽  
Fibrillin 1 ◽  
Healthy Control

Marfan syndrome is one of the most common dominantly inherited connective tissue disorders, affecting 2–3 in 10,000 individuals, and is caused by one of over 2800 unique FBN1 mutations. Mutations in FBN1 result in reduced fibrillin-1 expression, or the production of two different fibrillin-1 monomers unable to interact to form functional microfibrils. Here, we describe in vitro evaluation of antisense oligonucleotides designed to mediate exclusion of FBN1 exon 52 during pre-mRNA splicing to restore monomer homology. Antisense oligonucleotide sequences were screened in healthy control fibroblasts. The most effective sequence was synthesised as a phosphorodiamidate morpholino oligomer, a chemistry shown to be safe and effective clinically. We show that exon 52 can be excluded in up to 100% of FBN1 transcripts in healthy control fibroblasts transfected with PMO52. Immunofluorescent staining revealed the loss of fibrillin 1 fibres with ~50% skipping and the subsequent re-appearance of fibres with >80% skipping. However, the effect of exon skipping on the function of the induced fibrillin-1 isoform remains to be explored. Therefore, these findings demonstrate proof-of-concept that exclusion of an exon from FBN1 pre-mRNA can result in internally truncated but identical monomers capable of forming fibres and lay a foundation for further investigation to determine the effect of exon skipping on fibrillin-1 function.

scholarly journals Superior Silencing by 2′,4′-BNANC-Based Short Antisense Oligonucleotides Compared to 2′,4′-BNA/LNA-Based Apolipoprotein B Antisense Inhibitors

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Tsuyoshi Yamamoto ◽  
Hidenori Yasuhara ◽  
Fumito Wada ◽  
Mariko Harada-Shiba ◽  
Takeshi Imanishi ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Inhibitory Activity ◽  
Apolipoprotein B ◽  
Antisense Oligonucleotides ◽  
Cost Effective ◽  
Fine Tuning ◽  
Locked Nucleic Acid ◽  
Penalty Term ◽  
Target Mrna

The duplex stability with target mRNA and the gene silencing potential of a novel bridged nucleic acid analogue are described. The analogue,2′,4′-BNANCantisense oligonucleotides (AONs) ranging from 10- to 20-nt-long, targeted apolipoprotein B.2′,4′-BNANCwas directly compared to its conventional bridged (or locked) nucleic acid (2′,4′-BNA/LNA)-based counterparts. Melting temperatures of duplexes formed between2′,4′-BNANC-based antisense oligonucleotides and the target mRNA surpassed those of 2′,4′-BNA/LNA-based counterparts at all lengths. Anin vitrotransfection study revealed that when compared to the identical length2′,4′-BNA/LNA-based counterpart, the corresponding2′,4′-BNANC-based antisense oligonucleotide showed significantly stronger inhibitory activity. This inhibitory activity was more pronounced in shorter (13-, 14-, and 16-mer) oligonucleotides. On the other hand, the 2′,4′-BNANC-based 20-mer AON exhibited the highest affinity but the worstIC50value, indicating that very high affinity may undermine antisense potency. These results suggest that the potency of AONs requires a balance between reward term and penalty term. Balance of these two parameters would depend on affinity, length, and the specific chemistry of the AON, and fine-tuning of this balance could lead to improved potency. We demonstrate that2′,4′-BNANCmay be a better alternative to conventional2′,4′-BNA/LNA, even for “short” antisense oligonucleotides, which are attractive in terms of drug-likeness and cost-effective bulk production.

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