Publication: Consequences of Making the Inactive Active Through Changes in Antisense Oligonucleotide Chemistries
Issued Date
2019-12-20
Resource Type
ISSN
16648021
Other identifier(s)
2-s2.0-85077442171
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Frontiers in Genetics. Vol.10, (2019)
Suggested Citation
Khine Zaw, Kane Greer, May Thandar Aung-Htut, Chalermchai Mitrpant, Rakesh N. Veedu, Sue Fletcher, Steve D. Wilton Consequences of Making the Inactive Active Through Changes in Antisense Oligonucleotide Chemistries. Frontiers in Genetics. Vol.10, (2019). doi:10.3389/fgene.2019.01249 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/49992
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Consequences of Making the Inactive Active Through Changes in Antisense Oligonucleotide Chemistries
Abstract
© Copyright © 2019 Zaw, Greer, Aung-Htut, Mitrpant, Veedu, Fletcher and Wilton. Antisense oligonucleotides are short, single-stranded nucleic acid analogues that can interfere with pre-messenger RNA (pre-mRNA) processing and induce excision of a targeted exon from the mature transcript. When developing a panel of antisense oligonucleotides to skip every dystrophin exon, we found great variation in splice switching efficiencies, with some antisense oligonucleotides ineffective, even when directed to canonical splice sites and transfected into cells at high concentrations. In this study, we re-evaluated some of these ineffective antisense oligonucleotide sequences after incorporation of locked nucleic acid residues to increase annealing potential. Antisense oligonucleotides targeting exons 16, 23, and 51 of human DMD transcripts were synthesized as two different chemistries, 2′-O-methyl modified bases on a phosphorothioate backbone or mixmers containing several locked nucleic acid residues, which were then transfected into primary human myotubes, and DMD transcripts were analyzed for exon skipping. The ineffective 2′-O-methyl modified antisense oligonucleotides induced no detectable exon skipping, while all corresponding mixmers did induce excision of the targeted exons. Interestingly, the mixmer targeting exon 51 induced two unexpected transcripts arising from partial skipping of exon 51 with retention of 95 or 188 bases from the 5′ region of exon 51. These results indicated that locked nucleic acid/2′-O-methyl mixmers are more effective at inducing exon skipping, however, this improvement may come at the cost of activating alternative cryptic splice sites and off-target effects on gene expression.