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.

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 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.

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.

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

Distinguishing L from M photopigment coding sequences by hybridization to novel locked nucleic acid (LNA) oligonucleotide probes

2008 ◽  
Vol 25 (3) ◽  
pp. 283-287
Author(s):  
CHRISTINA PETTAN-BREWER ◽  
LI FU ◽  
SAMIR S. DEEB
Keyword(s):  
Nucleic Acid ◽  
Locked Nucleic Acid ◽  
Macaca Nemestrina ◽  
Oligonucleotide Probes ◽  
Coding Sequences ◽  
Cone Mosaic ◽  
Target Nucleic Acid ◽  
High Degree

Many attempts have been made over the years to distinguish human and primate L (long-wavelength sensitive) from M (middle-wavelength sensitive) cone photoreceptors using either immunohistochemistry or in situ hybridization. These attempts have been unsuccessful due to the very high degree of identity between the sequences of the L and M proteins and encoding mRNAs. The recent development of chemically modified oligonucleotide probes, referred to as locked nucleic acid (LNA) probes, has shown that they hybridize with much greater affinity and specificity to the target nucleic acid. This has greatly increased the potential for differentiating L from M cones by in situ hybridization. We have designed LNA oligonucleotide probes that are complementary to either the L or M coding sequences located in exon 5 of the Macaca nemestrina L and M pigment genes. We have shown that the LNA-M and LNA-L probes hybridize specifically to their respective target nucleic acid sequences in vitro. This result strongly suggests that these probes would be instrumental in rapidly distinguishing L from M cone in the entire retina, and in defining the cone mosaic during development and in adults.

Bioreducible poly(urethane amine)s for robust nucleic acid transfection in stem cells

2019 ◽  
Vol 7 (8) ◽  
pp. 3510-3518
Author(s):  
Ying Ye ◽  
Rong Jin ◽  
Xiaoxin Hu ◽  
Juhua Zhuang ◽  
Wei Xia ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Nucleic Acid ◽  
Embryonic Stem ◽  
Highly Efficient ◽  
Efficient Delivery ◽  
Low Cytotoxicity

Bioreducible poly(urethane amine)s can be designed and optimized for highly efficient delivery in vitro of DNA or SiRNA into adult or embryonic stem cells with low cytotoxicity.

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.

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