Nuclease-resistant composite 2',5'-oligoadenylate-3', 5'-oligonucleotides for the targeted destruction of RNA: 2-5A-iso-antisense.
A new modification of 2-5A-antisense, 2-5A-iso-antisense, has been developed based on a reversal of the direction of the polarity of the antisense domain of a 2-5A-antisense composite nucleic acid. This modification was able to anneal with its target RNA as well as the parental 2-5A-antisense chimera. The 2-5A-iso-antisense oligonucleotide displayed enhanced resistance to degradation by 3'-exonuclease enzyme activity such as that represented by snake venom phosphodiesterase and by that found in human serum. 2-5A-Iso-antisense was able to effect the degradation of a synthetic nontargeted substrate, [5'-32P]pC11U2C7, and two targeted RNAs, PKR and BCR mRNAs, in a cell-free system containing purified recombinant human 2-5A-dependent RNase L. These results demonstrated that the novel structural modification represented by 2-5A-iso-antisense provided a stabilized biologically active formulation of the 2-5A-antisense strategy.
Xiao
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W
Section on Biomedical Chemistry, Laboratory of Medicinal Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
Li
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G
Player
M R
MR
Maitra
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RK
Waller
C F
CF
Silverman
R H
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Torrence
P F
PF
D018076:DNA, Complementary / Q000378:metabolism; D004722:Endoribonucleases / Q000378:metabolism; D016044:Fusion Proteins, bcr-abl / Q000378:metabolism; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000378:metabolism; D009843:Oligoribonucleotides / Q000378:metabolism; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000097:blood / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000378:metabolism; D019892:eIF-2 Kinase / Q000378:metabolism
D018076:DNA, Complementary; D016376:Oligonucleotides, Antisense; D009843:Oligoribonucleotides; D012334:RNA, Neoplasm; D012313:RNA; D016044:Fusion Proteins, bcr-abl; D019892:eIF-2 Kinase; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I; C470790:phosphodiesterase I, snake venom