Inhibition of off-target cleavage by RNase H using an artificial cationic oligosaccharide

Hara, Rintaro Iwata; Wada, Takehiko

doi:10.1039/d1ob00983d

Cited by 4 publications

(2 citation statements)

References 31 publications

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“…This data demonstrates that optimization of ASO size and the number of LNA nt provides a tool for finding balanced ASO sequence with moderate affinity and moderate specificity but does not resolve the affinity/specificity dilemma 19 . ASO modified by other artificial nucleotides 55 or conjugated with ASO/RNA complex stabilizing groups 56 , 57 should experience the same fundamental challenge.…”

Section: Antisense Oligonucleotides (Aso Agents)mentioning

confidence: 99%

Specificity of oligonucleotide gene therapy (OGT) agents

Nedorezova¹,

Dubovichenko²,

Belyaeva³

et al. 2022

Theranostics

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Oligonucleotide gene therapy (OGT) agents (e. g. antisense, deoxyribozymes, siRNA and CRISPR/Cas) are promising therapeutic tools. Despite extensive efforts, only few OGT drugs have been approved for clinical use. Besides the problem of efficient delivery to targeted cells, hybridization specificity is a potential limitation of OGT agents. To ensure tight binding, a typical OGT agent hybridizes to the stretch of 15-25 nucleotides of a unique targeted sequence. However, hybrids of such lengths tolerate one or more mismatches under physiological conditions, the problem known as the affinity/specificity dilemma. Here, we assess the scale of this problem by analyzing OGT hybridization-dependent off-target effects (HD OTE) in vitro , in animal models and clinical studies. All OGT agents except deoxyribozymes exhibit HD OTE in vitro , with most thorough evidence of poor specificity reported for siRNA and CRISPR/Cas9. Notably, siRNA suppress non-targeted genes due to (1) the partial complementarity to mRNA 3'-untranslated regions (3'-UTR), and (2) the antisense activity of the sense strand. CRISPR/Cas9 system can cause hundreds of non-intended dsDNA breaks due to low specificity of the guide RNA, which can limit therapeutic applications of CRISPR/Cas9 by ex-vivo formats. Contribution of this effects to the observed in vivo toxicity of OGT agents is unclear and requires further investigation. Locked or peptide nucleic acids improve OGT nuclease resistance but not specificity. Approaches that use RNA marker dependent (conditional) activation of OGT agents may improve specificity but require additional validation in cell culture and in vivo .

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Section: Antisense Oligonucleotides (Aso Agents)mentioning

confidence: 99%

Specificity of oligonucleotide gene therapy (OGT) agents

Nedorezova¹,

Dubovichenko²,

Belyaeva³

et al. 2022

Theranostics

View full text Add to dashboard Cite

show abstract

“…17 Most recently, we have reported that ODAGal4 is effective for modulating the mismatch discrimination by the RNase H cleavage. 19 Moreover, ODAGal is useful for the selective delivery of a siRNA to the liver cell by conjugating vitamin E to ODAGal. 20 Notably, it has been suggested that the presence of ODAGals does not disturb the gene silencing effect of siRNAs.…”

Section: Introductionmentioning

confidence: 99%