Efficient RNase H-directed cleavage of RNA promoted by antisense DNA or 2'F-ANA constructs containing acyclic nucleotide inserts.
The ability of modified antisense oligonucleotides (AONs) containing acyclic interresidue units to support RNase H-promoted cleavage of complementary RNA is described. Manipulation of the backbone and sugar geometries in these conformationally labile monomers shows great benefits in the enzymatic recognition of the nucleic acid hybrids, while highlighting the importance of local strand conformation on the hydrolytic efficiency of the enzyme more conclusively. Our results demonstrate that the duplexes support remarkably high levels of enzymatic degradation when treated with human RNase HII, making them efficient mimics of the native substrates. Furthermore, interesting linker-dependent modulation of enzymatic activity is observed during in vitro assays, suggesting a potential role for this AON class in an RNase H-dependent pathway of controlling RNA expression. Additionally, the butyl-modified 2'F-ANA AONs described in this work constitute the first examples of a nucleic acid species capable of eliciting high RNase H activity while possessing a highly flexible molecular architecture at predetermined sites along the AON.
Mangos
Maria M
MM
Contribution from McGill University, Department of Chemistry, Montreal, QC, Canada.
Min
Kyung-Lyum
KL
Viazovkina
Ekaterina
E
Galarneau
Annie
A
Elzagheid
Mohamed I
MI
Parniak
Michael A
MA
Damha
Masad J
MJ
D001089:Arabinose / Q000031:analogs & derivatives* / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D016373:DNA, Antisense / Q000138:chemical synthesis / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology; D011905:Genes, ras; D006801:Humans; D009690:Nucleic Acid Conformation; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000737:chemistry*; D018075:RNA, Complementary / Q000737:chemistry* / Q000378:metabolism; D016914:Ribonuclease H / Q000737:chemistry* / Q000378:metabolism; D013329:Structure-Activity Relationship
C118537:2'-deoxy-2'-fluoroarabinose; D016373:DNA, Antisense; D009692:Nucleic Acid Heteroduplexes; D016376:Oligonucleotides, Antisense; D018075:RNA, Complementary; D004247:DNA; D001089:Arabinose; D016914:Ribonuclease H