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Propynylated phosphodiester oligonucleotides inhibit ICAM-1 expression in A549 cells on electroporation
L Meunier1, M Monsigny, A C Roche
1Glycobiologie, Vectorologie et Trafic Intracellulaire, Centre de Biophysique Moléculaire, CNRS, Orléans, France.
Abstract:
Oligodeoxynucleotides (ODN) are used largely as either primers, antisense, or triplex-forming units. Phosphodiester ODN (PO-ODN), which are very rapidly degraded by exonucleases, must be protected at their ends. Even so, their life span inside cells is quite short. Phosphorothioate ODN (PS-ODN) are less sensitive to nucleases and are extensively used as antisense. Unfortunately, unlike PO-ODN, they interact with a number of molecules, including proteins, in addition to their specific nucleic acid targets. Their affinity for their target is lower than that of PO-ODN. PS-ODN containing propyne groups on C5 of pyrimidine have been shown to have a higher affinity toward their nucleic acid target. Here, we show that propynylated PO-ODN are more stable and much more efficient than their propyne-free counterparts. They are not efficient when they are used as lipoplexes, but they act as specific antisense on electroporation.
Insights
Propynylated phosphodiester oligodeoxynucleotides (PO-ODN) show enhanced stability and efficiency compared to standard PO-ODN. These modified oligonucleotides effectively function as antisense agents when delivered via electroporation.
Area of Science:
- Oligonucleotide chemistry and molecular biology
Background:
- Oligodeoxynucleotides (ODN) are crucial in molecular biology, serving as primers, antisense agents, or triplex-forming units.
- Standard phosphodiester ODN (PO-ODN) exhibit short cellular lifespans due to rapid exonuclease degradation, necessitating end protection.
- Phosphorothioate ODN (PS-ODN) offer increased nuclease resistance but have lower target affinity and non-specific protein interactions compared to PO-ODN.
Purpose of the Study:
- To investigate the impact of propynylation on the stability and efficiency of PO-ODN.
- To evaluate the antisense activity of propynylated PO-ODN using different delivery methods.
Main Methods:
- Synthesis of propynylated PO-ODN with propyne groups on C5 of pyrimidines.
- Assessment of ODN stability against nucleases.
- Evaluation of antisense efficacy using lipoplex and electroporation delivery methods.
Main Results:
- Propynylated PO-ODN demonstrated significantly improved stability and efficiency over unmodified PO-ODN.
- Lipoplex delivery of propynylated PO-ODN resulted in low efficiency.
- Electroporation enabled specific antisense activity of propynylated PO-ODN.
Conclusions:
- Propynylation is a promising strategy to enhance the performance of PO-ODN.
- Electroporation is an effective delivery method for propynylated PO-ODN to achieve specific antisense effects.