Properties of base-pairing in the stem region of hairpin antisense oligonucleotides containing 2′-methoxynucleosides

Hosono, Kuniaki; Kuwasaki, Tomoyuki; Tsukahara, Satoru; Takai, Kazuyuki; Takaku, Hiroshi

doi:10.1016/0304-4165(95)00053-e

Cited by 2 publications

(2 citation statements)

References 13 publications

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“…The nuclease resistance of this miRNase is not inferior to the stability of potent phosphorothioate DNA/LNA mixmer, which begins to degrade by 24 h of similar incubation (Lennox and Behlke, 2010). The obtained results correlate with the data reported earlier, where it was shown that 3′-end structures can improve resistance to snake venom phosphodiesterase, DNA polymerase I, and FBS and significantly increase functional potency for target RNA knockdown (Tang et al, 1993; Hosono et al, 1995). The presence of flanking duplexes also improves the ability of anti-miRNA oligonucleotides to invade RISC and serve functional enhancement (Vermeulen et al, 2007; Lennox and Behlke, 2011).…”

Section: Discussionsupporting

confidence: 90%

Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model

et al. 2019

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Control of the expression of oncogenic small non-coding RNAs, notably microRNAs (miRNAs), is an attractive therapeutic approach. We report a design platform for catalytic knockdown of miRNA targets with artificial, sequence-specific ribonucleases. miRNases comprise a peptide [(LeuArg) 2 Gly] 2 capable of RNA cleavage conjugated to the miRNA-targeted oligodeoxyribonucleotide, which becomes nuclease-resistant within the conjugate design, without resort to chemically modified nucleotides. Our data presented here showed for the first time a truly catalytic character of our miR-21-miRNase and its ability to cleave miR-21 in a multiple catalytic turnover mode. We demonstrate that miRNase targeted to miR-21 (miR-21-miRNase) knocked down malignant behavior of tumor cells, including induction of apoptosis, inhibition of cell invasiveness, and retardation of tumor growth, which persisted on transplantation into mice of tumor cells treated once with miR-21-miRNase. Crucially, we discover that the high biological activity of miR-21-miRNase can be directly related not only to its truly catalytic sequence-specific cleavage of miRNA but also to its ability to recruit the non-sequence specific RNase H found in most cells to elevate catalytic turnover further. miR-21-miRNase worked synergistically even with low levels of RNase H. Estimated degradation in the presence of RNase H exceeded 10 3 miRNA target molecules per hour for each miR-21-miRNase molecule, which provides the potency to minimize delivery requirements to a few molecules per cell. In contrast to the comparatively high doses required for the simple steric block of antisense oligonucleotides, truly catalytic inactivation of miRNA offers more effective, irreversible, and persistent suppression of many copy target sequences. miRNase design can be readily adapted to target other pathogenic microRNAs overexpressed in many disease states.

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Section: Discussionsupporting

confidence: 90%

Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model

et al. 2019

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“…However, when the flanking sequence on both ends was duplexed with short complementary 2′OMe oligonucleotides or the flanking sequences formed hairpin structures, potency was markedly increased. There is precedent from previous antisense studies demonstrating that 3′-end hairpin structures can improve nuclease stability and significantly increase functional potency for knockdown of RNA targets (48,49). It was also suggested that the presence of flanking duplex domains improved the ability of the AMO to invade RISC and served a functional purpose beyond nuclease stabilization (35).…”

Section: Direct Comparison Of Amo Potenciesmentioning

confidence: 95%