Titel
mCherry on Top: A Positive Read-Out Cellular Platform for Screening DMD Exon Skipping Xenopeptide–PMO Conjugates
Autor*in
Anna-Lina Lessl
Pharmaceutical Biotechnology, Department of Pharmacy, LMU Munich
Autor*in
Jana Pöhmerer
Pharmaceutical Biotechnology, Department of Pharmacy, LMU Munich
Autor*in
Yi Lin
Pharmaceutical Biotechnology, Department of Pharmacy, LMU Munich
... show all
Abstract
Phosphorodiamidate morpholino oligomers (PMOs) are a special type of antisense oligonucleotides (ASOs) that can be used as therapeutic modulators of pre-mRNA splicing. Application of nucleic-acid-based therapeutics generally requires suitable delivery systems to enable efficient transport to intended tissues and intracellular targets. To identify potent formulations of PMOs, we established a new in vitro–in vivo screening platform based on mdx exon 23 skipping. Here, a new in vitro positive read-out system (mCherry-DMDEx23) is presented that is sensitive toward the PMO(Ex23) sequence mediating DMD exon 23 skipping and, in this model, functional mCherry expression. After establishment of the reporter system in HeLa cells, a set of amphiphilic, ionizable xenopeptides (XPs) was screened in order to identify potent carriers for PMO delivery. The identified best-performing PMO formulation with high splice-switching activity at nanomolar concentrations in vitro was then translated to in vivo trials, where exon 23 skipping in different organs of healthy BALB/c mice was confirmed. The predesigned in vitro–in vivo workflow enables evaluation of PMO(Ex23) carriers without change of the PMO sequence and formulation composition. Furthermore, the identified PMO–XP conjugate formulation was found to induce highly potent exon skipping in vitro and redistributed PMO activity in different organs in vivo.
Stichwort
AnatomyConjugate acid-base pairsFlow cytometryGeneticsRodent models
Objekt-Typ
Sprache
Englisch [eng]
Erschienen in
Titel
Bioconjugate Chemistry
Band
34
Ausgabe
12
ISSN
1043-1802
Erscheinungsdatum
2023
Seitenanfang
2263
Seitenende
2274
Publication
American Chemical Society (ACS)
Erscheinungsdatum
2023
Zugänglichkeit
Rechteangabe
© 2023 The Authors

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