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CC BY-NC V4.0, Open Access
Journal article
Enhancing the intracellular delivery of antisense oligonucleotides (ASO) : a comparative study of aptamer, vitamin E, and cholesterol ASO conjugates
RSC advances, Vol.15(51), pp.43727-43736
2025
PMID: 41220634
Abstract
Antisense oligonucleotides (ASOs) have emerged as powerful tools for gene modulation; however, their clinical application is often hindered by inefficient intracellular delivery. Recent advances suggest that conjugation to biomolecules such as aptamers, lipids, or vitamins may enhance uptake and efficacy. This study investigates the potential of aptamer, vitamin E, and cholesterol-conjugated ASOs to improve delivery and functional activity in cancer cell models. The ASO PNAT524 was conjugated to two DNA aptamers-AS1411 and S2.2-via thiol and triethylene glycol (TEG) linkers, respectively. In parallel, PNAT524 was modified with vitamin E and cholesterol moieties. The conjugates were evaluated for cellular uptake, exon-skipping activity, and cytotoxicity in cancer cell lines. Fluorescence microscopy was used to determine subcellular localization. Aptamer conjugation (AS1411, S2.2) did not significantly enhance exon-skipping efficiency compared to unconjugated PNAT524, consistent with previous findings. In contrast, vitamin E and cholesterol conjugates demonstrated potent, dose-dependent exon-skipping activity and cytotoxic effects. Among all formulations, the cholesterol-conjugated ASO (524-Chol) showed the highest efficacy, with superior splice-modulating and cytotoxic outcomes. Fluorescence microscopy confirmed nuclear and cytoplasmic localization of lipid-conjugated ASOs. These findings indicate that aptamer conjugation provides minimal benefit for ASO delivery, while cholesterol and vitamin E conjugation significantly enhance intracellular delivery and therapeutic activity. The 524-Chol conjugate holds strong potential for adaptation in ASOs targeting EGFR and other oncogenes, representing a promising avenue for ASO-based cancer therapeutics.
Details
- Title
- Enhancing the intracellular delivery of antisense oligonucleotides (ASO) : a comparative study of aptamer, vitamin E, and cholesterol ASO conjugates
- Authors/Creators
- Akilandeswari Ashwini Balachandran - Perron Institute for Neurological and Translational ScienceBal Hari Poudel - Murdoch University, Personalised Medicine CentreKamal Rahimizadeh - Precision Nucleic Acid Therapeutics, Perron Institute for Neurological and Translational Science Nedlands Western Australia 6009 AustraliaArpitha Chikkanna - Perron Institute for Neurological and Translational ScienceRakesh N Veedu - Precision Nucleic Acid Therapeutics, Perron Institute for Neurological and Translational Science Nedlands Western Australia 6009 Australia
- Publication Details
- RSC advances, Vol.15(51), pp.43727-43736
- Publisher
- Royal Society of Chemistry
- Number of pages
- 10
- Identifiers
- 991005830347207891
- Copyright
- © 2025 The Author(s).
- Murdoch Affiliation
- Personalised Medicine Centre
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- Citation topics
- 2 Chemistry
- 2.170 Nucleic Acids Chemistry
- 2.170.988 Oligonucleotide Modifications
- Web Of Science research areas
- Chemistry, Multidisciplinary
- ESI research areas
- Chemistry