Bulge-Forming miRNases Cleave Oncogenic miRNAs at the Central Loop Region in a Sequence-Specific Manner

International Journal of Molecular Sciences

Bahareh Amirloo¹  Elena Bichenkova¹  David Clarke¹  Daria Chiglintseva²  Svetlana Gaponova²  Olga Patutina²  Valentin Vlassov²  Marina Zenkova² 

[1] Faculty of Biology, Medicine and Health, School of Health Sciences, University of Manchester, Oxford Rd, Manchester M13 9PT, UK;Institute of Chemical Biology and Fundamental Medicine SB RAS, Lavrentiev’s Ave. 8, 630090 Novosibirsk, Russia;

关键词: microRNA;    miRNases;    peptide–oligonucleotide conjugates;    artificial ribonucleases;    hybridization;    RNA cleavage;   

DOI  :  10.3390/ijms23126562

来源: DOAJ

【 摘 要 】

The selective degradation of disease-associated microRNA is promising for the development of new therapeutic approaches. In this study, we engineered a series of bulge-loop-forming oligonucleotides conjugated with catalytic peptide [(LeuArg)₂Gly]₂ (BC–miRNases) capable of recognizing and destroying oncogenic miR-17 and miR-21. The principle behind the design of BC–miRNase is the cleavage of miRNA at a three-nucleotide bulge loop that forms in the central loop region, which is essential for the biological competence of miRNA. A thorough study of mono- and bis-BC–miRNases (containing one or two catalytic peptides, respectively) revealed that: (i) the sequence of miRNA bulge loops and neighbouring motifs are of fundamental importance for efficient miRNA cleavage (i.e., motifs containing repeating pyrimidine–A bonds are more susceptible to cleavage); (ii) the incorporation of the second catalytic peptide in the same molecular scaffold increases the potency of BC–miRNase, providing a complete degradation of miR-17 within 72 h; (iii) the synergetic co-operation of BC–miRNases with RNase H accelerates the rate of miRNA catalytic cleavage by both the conjugate and the enzyme. Such synergy allows the rapid destruction of constantly emerging miRNA to maintain sufficient knockdown and achieve a desired therapeutic effect.

【 授权许可】

Unknown