title: Synthesis of a Morpholino Nucleic Acid (MNA)-Uridine Phosphoramidite, and Exon Skipping Using MNA/2'-O-Methyl Mixmer Antisense Oligonucleotide.
abstract: In this study, we synthesised a morpholino nucleoside-uridine (MNA-U) phosphoramidite and evaluated the potential of a MNA-modified antisense oligonucleotide (AO) sequences to induce exon 23 skipping inmdxmouse myotubes in vitro towards extending the applicability of morpholino chemistry with other nucleotide monomers. We designed, synthesised, and compared exon skipping efficiencies of 20 mer MNA-modified 2'-O-methyl RNA mixmer AO on a phosphorothioate backbone (MNA/2'-OMePS) to the corresponding fully modified 2'-O-methyl RNA AO (2'-OMePS) as a control. Our results showed that the MNA/2'-OMePS efficiently induced exon 23 skipping. As expected, the 2'-OMePS AO control yielded efficient exon 23 skipping. Under the applied conditions, both the AOs showed minor products corresponding to exon 22/23 dual exon skipping in low yield. As these are very preliminary data, more detailed studies are necessary; however, based on the preliminary results, MNA nucleotides might be useful in constructing antisense oligonucleotides.
authors.affiliation: Centre for Comparative Genomics, Murdoch University, Perth 6150, Australia. S.Chen@murdoch.edu.au.|Western Australian Neuroscience Research Institute, Perth 6150, Australia. S.Chen@murdoch.edu.au.
authors.forename: Suxiang
authors.initials: S
authors.lastname: Chen
authors.affiliation: Centre for Comparative Genomics, Murdoch University, Perth 6150, Australia. T.Le2@murdoch.edu.au.|Western Australian Neuroscience Research Institute, Perth 6150, Australia. T.Le2@murdoch.edu.au.
authors.forename: Bao T
authors.initials: BT
authors.lastname: Le
authors.affiliation: Centre for Comparative Genomics, Murdoch University, Perth 6150, Australia. rahimizadeh.kamal@gmail.com.
authors.forename: Kamal
authors.initials: K
authors.lastname: Rahimizadeh
authors.affiliation: GMK Research Laboratories Pvt. Ltd., Mallapur, Hyderabad 500 076, India. khalil.shaikh@gmkresearch.com.
authors.forename: Khalil
authors.initials: K
authors.lastname: Shaikh
authors.affiliation: GMK Research Laboratories Pvt. Ltd., Mallapur, Hyderabad 500 076, India. narinder.mohal@gmkresearch.com.
authors.forename: Narinder
authors.initials: N
authors.lastname: Mohal
authors.affiliation: Centre for Comparative Genomics, Murdoch University, Perth 6150, Australia. rveedu@ccg.murdoch.edu.au.|Western Australian Neuroscience Research Institute, Perth 6150, Australia. rveedu@ccg.murdoch.edu.au.
authors.forename: Rakesh N
authors.initials: RN
authors.lastname: Veedu
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016189:Dystrophin / Q000235:genetics*; D005091:Exons; D051379:Mice; D015394:Molecular Structure; D060172:Morpholinos / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects*; D009943:Organophosphorus Compounds / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D014529:Uridine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology
chemical_list: D016189:Dystrophin; D060172:Morpholinos; D009943:Organophosphorus Compounds; C085339:apo-dystrophin 1; C101323:uridine phosphoramidite; D014529:Uridine
keywords: PMO; antisense oligonucleotide; exon skipping; morpholino nucleotide