A novel photoelectrochemical strategy for sequence-spot bispecific analysis of N6-methyladenosine modification based on proximity ligation-triggered cascade amplification

• Published in: Analytica chimica acta 2023-07, Vol.1265, p.341287-341287, Article 341287
• Main Authors: Li, Yan, Sun, Yumeng, Yang, Weiguo, Yang, Lan, Su, Mingli, Fang, Lichao, Zheng, Junsong, Yuan, Ruo, Liang, Wenbin
• Format: Article
• Language: English
• Subjects: Amplification; Bioassay; N6-methyladenosine; Photoelectrochemical strategy; Proximity ligation assay
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2023.341287

N6-methyladenosine (m6A) modification as the most prevalent mammalian RNA internal modification has been considered as the invasive biomarkers in clinical diagnosis and biological mechanism researches. It is still challenged to explore m6A functions due to technical limitations on base- and location-resolved m6A modification. Herein, we firstly proposed a sequence-spot bispecific photoelectrochemical (PEC) strategy based on in situ hybridization mediated proximity ligation assay for m6A RNA characterization with high sensitivity and accuracy. Firstly, the target m6A methylated RNA could be transferred to the exposed cohesive terminus of H1 based on the special self-designed auxiliary proximity ligation assay (PLA) with sequence-spot bispecific recognition. The exposed cohesive terminus of H1 could furtherly trigger the next catalytic hairpin assembly (CHA) amplification and in situ exponential nonlinear hyperbranched hybridization chain reaction for highly sensitive monitoring of m6A methylated RNA. Compared with conventional technologies, the proposed sequence-spot bispecific PEC strategy for m6A methylation of special RNA based on proximity ligation-triggered in situ nHCR performed improved sensitivity and selectivity with a detection limit of 53 fM, providing new insights into highly sensitive monitoring m6A methylation of RNA in bioassay, disease diagnosis and RNA mechanism. [Display omitted] •Sequence-Site bispecific recognition provide a more accurate and sensitive way to detect N6-methyladenosine modification.•Photoelectrochemistry performed with in situ hybridization mediated proximity linkage analysis.•In situ nonlinear hyperbranched hybridization chain reaction realizes high signal amplification.