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Chemically Modified Oligonucleotide–Increased Stability Negatively Correlates with Its Efficacy Despite Efficient Electrotransfer

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Abstract

Despite great potential for disease treatment, small interfering RNA (siRNA) development has been hampered due to its poor stability and the lack of efficient delivery method. To overcome the sensitivity, new generations of chemically modified oligonucleotides have been developed such as the locked nucleic acid (LNA). LNA substitution in an siRNA sequence (siLNA) is supposed to increase its stability and its affinity for its complementary sequence. The purpose of this study was to evaluate the potential benefit of an anti-GFP siLNA using the biophysical delivery method electropermeabilization. We used two types of electrical conditions: electrochemotherapy (ECT), a condition for efficient transfer of small molecules in clinics, and electrogenotherapy (EGT), a condition for efficient transfer of macromolecules. We first confirmed that siLNA was indeed more stable in mouse serum than unmodified siRNA. After determining the ECT and EGT optimal electrical parameters for a human colorectal carcinoma cell line (HCT-116) expressing eGFP, we showed that modifications of siRNA do not interfere with electrotransfer efficiency. However, despite its higher stability and its high electrotransfer efficacy, siLNA was less efficient for eGFP silencing compared to the electrotransferred, unmodified siRNA regardless of the electrical conditions used. Our study highlighted the care that is needed when designing chemically modified oligonucleotides.

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Acknowledgement

This work was supported by grants of the Ligue contre le Cancer, CNRS and FP7 Oncomirs (Grant 201102). Flow cytometry was performed at the TRI (funded by the region Midi-Pyrénées, the communauté du Grand Toulouse and the FEDER) platform at the IPBS. We thank Bettina Couderc (Institut National de la Santé et de la Recherche Médicale U563, Toulouse) for providing the HCT-GFP cell lines.

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Authors and Affiliations

  1. CNRS (Centre National de la Recherche Scientifique), IPBS (Institut de Pharmacologie et de Biologie Structurale), BP 64182, 205 route de Narbonne, 31077, Toulouse, France

    Sandrine Pelofy, Justin Teissié, Muriel Golzio & Sophie Chabot

  2. IPBS, Université Paul Sabatier de Toulouse, 31077, Toulouse, France

    Sandrine Pelofy, Justin Teissié, Muriel Golzio & Sophie Chabot

Authors
  1. Sandrine Pelofy
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  2. Justin Teissié
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  3. Muriel Golzio
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  4. Sophie Chabot
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Corresponding author

Correspondence to Muriel Golzio.

Additional information

M. Golzio and S. Chabot contributed equally to this study.

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Pelofy, S., Teissié, J., Golzio, M. et al. Chemically Modified Oligonucleotide–Increased Stability Negatively Correlates with Its Efficacy Despite Efficient Electrotransfer. J Membrane Biol 245, 565–571 (2012). https://doi.org/10.1007/s00232-012-9468-9

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  • DOI: https://doi.org/10.1007/s00232-012-9468-9

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