Highly efficient silencing of microRNA by heteroduplex oligonucleotides

Yoshioka, Koshiro; Kunieda, Takehisa; Asami, Yasushi; Guo, Huijia; Miyata, Haruka; Yoshida-Tanaka, Kie; Sujino, Yumiko; Piao, Wenying; Kuwahara, Hiroya; Nishina, Kazutaka; Hara, Rintaro Iwata; Nagata, Tetsuya; Wada, Takehiko; Obika, Satoshi; Yokota, Takanori

doi:10.1093/nar/gkz492

Cited by 37 publications

(33 citation statements)

References 63 publications

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“…The Toc-HDO (coRNA) design significantly enhanced the antisense potency of the parent ASO in mouse liver. [13][14][15][16][17] We also showed that the coRNA of the Toc-HDO (coRNA) is stable in the serum and that conjugation with Toc enhances productive delivery to the liver by enhancing association with serum lipoproteins. Toc-HDO (coRNA) was taken up into cells mostly in the intact form, after which the coRNA strand was cleaved by RNase H with the release of the parent ASO inside the cells.…”

Section: Introductionmentioning

confidence: 92%

Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo

et al. 2021

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We recently reported the antisense properties of a DNA/RNA heteroduplex oligonucleotide consisting of a phosphorothioate DNA-gapmer antisense oligonucleotide (ASO) strand and its complementary phosphodiester RNA/phosphorothioate 2 0 -Omethyl RNA strand. When a-tocopherol was conjugated with the complementary strand, the heteroduplex oligonucleotide silenced the target RNA more efficiently in vivo than did the parent single-stranded ASO. In this study, we designed a new type of the heteroduplex oligonucleotide, in which the RNA portion of the complementary strand was replaced with phosphodiester DNA, yielding an ASO/DNA double-stranded structure. The ASO/DNA heteroduplex oligonucleotide showed similar activity and liver accumulation as did the original ASO/RNA design. Structure-activity relationship studies of the complementary DNA showed that optimal increases in the potency and the accumulation were seen when the flanks of the phosphodiester DNA complement were protected using 2 0 -O-methyl RNA and phosphorothioate modifications. Furthermore, evaluation of the degradation kinetics of the complementary strands revealed that the DNA-complementary strand as well as the RNA strand were completely cleaved in vivo. Our results expand the repertoire of chemical modifications that can be used with the heteroduplex oligonucleotide technology, providing greater design flexibility for future therapeutic applications.

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Section: Introductionmentioning

confidence: 92%

Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo

et al. 2021

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“… 5 MicroRNA-targeting HDO (HDO-antimiR) displayed high potency in cleaving the target microRNA, whereas the parental ASO exerted its potency via a steric blocking (not cleaving) mechanism. 6 Additionally, HDO-antimiR conjugated with GalNAc was more potent in the liver than the parent ASO conjugated with GalNAc, where delivery efficiency of HDO was comparable to that of ASO. 6 These findings suggested that HDO may have a distinct intracellular trafficking pathway and processing machinery different from the single-stranded ASO.…”

Section: Introductionmentioning

confidence: 99%

“… 6 Additionally, HDO-antimiR conjugated with GalNAc was more potent in the liver than the parent ASO conjugated with GalNAc, where delivery efficiency of HDO was comparable to that of ASO. 6 These findings suggested that HDO may have a distinct intracellular trafficking pathway and processing machinery different from the single-stranded ASO.…”

Section: Introductionmentioning

confidence: 99%

“…We recently developed a novel highly efficient oligonucleotide, DNA/RNA heteroduplex oligonucleotide (HDO), which is composed of an antisense gapmer (DNA nucleotides flanked by LNAs) and complementary RNA (cRNA). 5 , 6 , 7 HDO showed improved delivery to the target tissue by conjugating tocopherol to the cRNA strand. Additionally, Toc-HDO presented 4.8 times higher gene silencing effect than parental ASO, revealed after measuring the delivered oligonucleotide content.…”

Section: Introductionmentioning

confidence: 99%

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Separation-related rapid nuclear transport of DNA/RNA heteroduplex oligonucleotide: unveiling distinctive intracellular trafficking

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Asada

Yui

et al. 2021

Molecular Therapy - Nucleic Acids

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DNA/RNA heteroduplex oligonucleotide (HDO), composed of DNA/locked nucleic acid (LNA) antisense oligonucleotide (ASO) and complementary RNA, is a next-generation antisense therapeutic agent. HDO is superior to the parental ASO in delivering to target tissues, and it exerts a more potent gene-silencing effect. In this study, we aimed to elucidate the intracellular trafficking mechanism of HDO-dependent gene silencing. HDO was more preferably transferred to the nucleus after transfection compared to the parental ASO. To determine when and where HDO is separated into the antisense strand (AS) and complementary strand (CS), we performed live-cell time-lapse imaging and fluorescence resonance energy transfer (FRET) assays. These assays demonstrated that HDO had a different intracellular trafficking mechanism than ASO. After endocytosis, HDO was separated in the early endosomes, and both AS and CS were released into the cytosol. AS was more efficiently transported to the nucleus than CS. Separation, endosomal release, and initiation of nuclear transport were a series of time-locked events occurring at a median of 30 s. CS cleavage was associated with efficient nuclear distribution and gene silencing in the nucleus. Understanding the unique intracellular silencing mechanisms of HDO will help us design more efficient drugs and might also provide insight into innate DNA/RNA cellular biology.

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“…Further studies have established the utilization of ASOs for the targeting of microRNAs, and ultimately an efficient approach to enhance protein production (Fig. 1 ) [ 65 , 79 , 91 , 92 ]. The microRNAs (short RNAs consisting of 21-23 nucleotides) inhibit the gene expression or protein translation and control associated gene networks.…”

Section: Introductionmentioning

confidence: 99%

RNA-based therapies: A cog in the wheel of lung cancer defense

et al. 2021

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Lung cancer (LC) is a heterogeneous disease consisting mainly of two subtypes, non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC), and remains the leading cause of death worldwide. Despite recent advances in therapies, the overall 5-year survival rate of LC remains less than 20%. The efficacy of current therapeutic approaches is compromised by inherent or acquired drug-resistance and severe off-target effects. Therefore, the identification and development of innovative and effective therapeutic approaches are critically desired for LC. The development of RNA-mediated gene inhibition technologies was a turning point in the field of RNA biology. The critical regulatory role of different RNAs in multiple cancer pathways makes them a rich source of targets and innovative tools for developing anticancer therapies. The identification of antisense sequences, short interfering RNAs (siRNAs), microRNAs (miRNAs or miRs), anti-miRs, and mRNA-based platforms holds great promise in preclinical and early clinical evaluation against LC. In the last decade, RNA-based therapies have substantially expanded and tested in clinical trials for multiple malignancies, including LC. This article describes the current understanding of various aspects of RNA-based therapeutics, including modern platforms, modifications, and combinations with chemo-/immunotherapies that have translational potential for LC therapies.

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Highly efficient silencing of microRNA by heteroduplex oligonucleotides

Cited by 37 publications

References 63 publications

Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo

Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo

Separation-related rapid nuclear transport of DNA/RNA heteroduplex oligonucleotide: unveiling distinctive intracellular trafficking

RNA-based therapies: A cog in the wheel of lung cancer defense

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