Utility of oligonucleotide in upregulating circular RNA production in a cellular model

• Published in: Scientific reports Vol. 14; no. 1; p. 8096
• Main Authors: Ni, Lu, Yamada, Takeshi, Nakatani, Kazuhiko
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 06.04.2024; Nature Publishing Group; Nature Portfolio
• Subjects: 639/638/92/500; 639/638/92/610; Circular RNA; Design; Disease; Exons; HeLa Cells; Humanities and Social Sciences; Humans; Introns; mRNA; multidisciplinary; Oligonucleotides; RNA - genetics; RNA - metabolism; RNA Precursors - metabolism; RNA Splicing - genetics; RNA, Circular - genetics; Science; Science (multidisciplinary); Spliceosomes; Splicing
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-58663-x

Circular RNAs (circRNAs), are a covalently closed, single-stranded RNA without 5′- and 3′-termini, commonly stem from the exons of precursor mRNAs (pre-mRNAs). They have recently garnered interest, with studies uncovering their pivotal roles in regulating various aspects of cell functions and disease progressions. A notable feature of circRNA lies in the mechanism of its biogenesis involving a specialized form of splicing: back-splicing. A splicing process that relies on interactions between introns flanking the circularizing exon to bring the up and downstream splice sites in proximity through the formation of a prerequisite hairpin structure, allowing the spliceosomes to join the two splice sites together to produce a circular RNA molecule. Based on this mechanism, we explored the feasibility of facilitating the formation of such a prerequisite hairpin structure by utilizing a newly designed oligonucleotide, CircuLarIzation Promoting OligoNucleotide ( CLIP-ON ), to promote the production of circRNA in cells. CLIP-ON was designed to hybridize with and physically bridge two distal sequences in the flanking introns of the circularizing exons. The feasibility of CLIP-ON was confirmed in HeLa cells using a model pre-mRNA, demonstrating the applicability of CLIP-ON as a trans-acting modulator to upregulate the production of circRNAs in a cellular environment.