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© Research
Publication : Antisense Research and Development

New Insights into the Resistance of α-Oligonucleotides to Nucleases

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Antisense Research and Development - 01 Jan 1994

Sophie Vichier-Guerre, Alain Pompon, Isabelle Lefebvre, Jean-Louis Imbach

Link to Pubmed [PMID] – 8061517

Antisense Res Dev. 1994 Spring;4(1):9-18

Several studies have shown that a-oligonucleotides (a-ONs) are more resistant to degradation by nucleases than are ß-oligonucleotides (ß-ONs), but a few exceptions have been reported. The present work indicates that the resistance of a-ONs to 3-exonucleases contained in calf or human sera strongly depends on their 3′- terminal sequence. When the 2 last residues were …A-G, …C-A, or …C-T, the degradation rates in tissue culture media with fetal calf serum were similar to those observed for ß-ONs, but stabilization factors up to 200 were observed when the 2 last residues were …T-C, …A-C, or …C-C, and intermediate stabilization factors were found with …G—A, …T-A, or …T-T terminal sequences. Other data confirm that a-ONs are significantly more resistant than ß-ONs to purified 5-exo- and endonucleases as well as to 3-exonucleases contained in snake venom or CEM cell lysates, but suggest that sequence specificity may also exist in these media. These findings, which are consistent with literature data and explain the behavior of the aforementioned exceptions, also emphasize that the stability observed in the presence of purified nucleases cannot be extrapolated to sera. Other results show that handling, heat inactivation, and storage conditions have little effect on the 3′- exonuclease activity of serum-containing media, but can completely modify the nuclease activities of cell extracts. This study, which was carried out by means of the accurate “on-line ISRP cleaning” HPLC technique, brings new insights to the general problem of oligonucleotide stability.

http://online.liebertpub.com/doi/abs/10.1089/ard.1994.4.9