Abasic pivot substitution harnesses target specificity of RNA interference

Lee, Hye Sook; Seok, Hee Young; Lee, Dong Ha; Ham, Juyoung; Lee, Wooje; Youm, Emilia Moonkyung; Yoo, Jin S eon; Lee, Yong Seung; Jang, Eun Sook; Wook, Sung

doi:10.1038/ncomms10154

Cited by 41 publications

(45 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It has only been reported as a target gene of two miRNA (miR-27a and miR-124) [22, 23]. However, relationship between miR-224 and PCSK9 is still unknown.…”

Section: Discussionmentioning

confidence: 99%

A retrospective study of NENs and miR-224 promotes apoptosis of BON-1 cells by targeting PCSK9 inhibition

Bai

Hua

et al. 2016

Oncotarget

View full text Add to dashboard Cite

Neuroendocrine neoplasms (NENs) represent relatively rare tumors. The lack of diagnostic, therapeutic method and prognostic factors makes them a challenge to us. We retrospectively reviewed the data of 205 NENs patients among which 157 cases were followed-up. Proprotein convertase subtilisin/kexin 9 (PCSK9), a regulator of low density lipoprotein cholesterol (LDL-C), was confirmed as a target gene of microRNA-224. We found an increased incidence of NENs from 2012 to 2015. Women were usually diagnosed at earlier stages than men (P < 0.05). Tumor grading was associated with primary tumor site, especially esophagus and cardia NENs all at G3 (P <0.001). Age, tumor grading and LDL-C levels were independent risk factors of digestive NENs. Low LDL-C level was significantly correlated with survival rate and median overall survival (OS, P < 0.05). MicroRNA-224 agomir and PCSK9 siRNA could promote apoptosis and suppress proliferation, invasion of BON-1 cells (P < 0.05), but increase the level of glucocorticoid (GC, P < 0.05). Taken together, age, tumor grading and LDL-C level are independent risk factors of NENs. The miR-224/PCSK9/GC axis binds to tumorigenesis and prognosis of pancreatic NENs (p-NENs).

show abstract

“…It has only been reported as a target gene of two miRNA (miR-27a and miR-124) [22, 23]. However, relationship between miR-224 and PCSK9 is still unknown.…”

Section: Discussionmentioning

confidence: 99%

A retrospective study of NENs and miR-224 promotes apoptosis of BON-1 cells by targeting PCSK9 inhibition

Bai

Hua

et al. 2016

Oncotarget

View full text Add to dashboard Cite

show abstract

“…4 Thus, the development of modifications that avoid miRNA-like off-targeting is an active area of research. 5–7 Importantly, recent advances in structural studies of key protein–RNA complexes of the RNAi pathway have enabled structure-guided optimization of siRNA properties. 8,9 …”

mentioning

confidence: 99%

Structure-Guided Control of siRNA Off-Target Effects

et al. 2016

View full text Add to dashboard Cite

Short interfering RNAs (siRNAs) are promising therapeutics that make use of the RNA interference (RNAi) pathway, but liabilities arising from the native RNA structure necessitate chemical modification for drug development. Advances in the structural characterization of components of the human RNAi pathway have enabled structure-guided optimization of siRNA properties. Here we report the 2.3 Å resolution crystal structure of human Argonaute 2 (hAgo2), a key nuclease in the RNAi pathway, bound to an siRNA guide strand bearing an unnatural triazolyl nucleotide at position 1 (g1). Unlike natural nucleotides, this analogue inserts deeply into hAgo2’s central RNA binding cleft and thus is able to modulate pairing between guide and target RNAs. The affinity of the hAgo2–siRNA complex for a seed-only matched target was significantly reduced by the triazolyl modification, while the affinity for a fully matched target was unchanged. In addition, siRNA potency for off-target repression was reduced (4-fold increase in IC50) by the modification, while on-target knockdown was improved (2-fold reduction in IC50). Controlling siRNA on-target versus microRNA (miRNA)-like off-target potency by projection of substituent groups into the hAgo2 central cleft from g1 is a new approach to enhance siRNA selectivity with a strong structural rationale.

show abstract

“…11,12,19 Indeed, a recent report showed that introduction of an abasic residue at position 6 of the guide strand reduced miRNA-like off-target effects. 20 However, T M studies with model RNA duplexes indicated that 7-EAA triazole NEtPip does not substantially destabilize the duplex structure (ΔT M = 1.5 °C replacing one A-U pair with one 7-EAA triazole NEtPip-U base pair in a 12 bp duplex where a mismatch destabilizes this duplex ΔT M = 8.5 °C) and a crystal structure of a 7-EAA triazole NEtPip-containing RNA duplex showed typical A-form geometry. 15 Thus the influence of 7-EAA triazole NEtPip on miRNA-like off targeting appears to arise from effects not previously observed with sugar modified guide strands.…”

Section: Resultsmentioning

confidence: 99%

Controlling miRNA-like off-target effects of an siRNA with nucleobase modifications

et al. 2017

View full text Add to dashboard Cite

SiRNAs can cause unintended gene silencing due to miRNA-like effects because of the similarity in function of an siRNA guide strand and a miRNA. Here we evaluate the effect on miRNA-like off targeting of introducing the adenosine derivative 7-EAA and triazoles prepared from 7-EAA at different positions in an siRNA guide strand. We find that a sterically demanding triazole placed in the RNA duplex major groove at position six of the guide strand dramatically reduces miRNA-like off targeting potency. A high-resolution structure of an RNA duplex bearing a novel, major-groove localized triazole is reported, which suggests that modified triazoles could be disrupting the hAgo2-guide-target RNA ternary complex. Five different triazole modifications were tested at the guide strand 6-position for effects on on-target and miRNA-like off target knockdown potency. A 7-EAA triazole bearing a benzylamine substituent displayed on-target knockdown activity as potent as the native siRNA, while having an IC50 against a miRNA-like off target > 100-fold higher. Melting temperature studies revealed no obvious correlation between potency in knockdown assays and a modification’s effect on duplex stability. These results, along with known structures of hAgo2-guide-target ternary complexes, are used to rationalize the effect of 7-EAA triazoles on miRNA-like off target effects.

show abstract

Abasic pivot substitution harnesses target specificity of RNA interference

Cited by 41 publications

References 50 publications

A retrospective study of NENs and miR-224 promotes apoptosis of BON-1 cells by targeting PCSK9 inhibition

A retrospective study of NENs and miR-224 promotes apoptosis of BON-1 cells by targeting PCSK9 inhibition

Structure-Guided Control of siRNA Off-Target Effects

Controlling miRNA-like off-target effects of an siRNA with nucleobase modifications

Contact Info

Product

Resources

About