2'- O-Methyl molecular beacon: a promising molecular tool that permits elimination of sticky-end pairing and improvement of detection sensitivity

Jiafeng Gao; Yang Li; Wenqin Li; Chaofei Zeng; Fengna Xi; Jiahao Huang; Liang Cui

doi:10.1039/d0ra07341e

2'- O-Methyl molecular beacon: a promising molecular tool that permits elimination of sticky-end pairing and improvement of detection sensitivity

RSC Adv. 2020 Nov 14;10(68):41618-41624. doi: 10.1039/d0ra07341e. eCollection 2020 Nov 11.

Authors

Jiafeng Gao¹, Yang Li², Wenqin Li², Chaofei Zeng¹, Fengna Xi¹, Jiahao Huang², Liang Cui¹

Affiliations

¹ Department of Chemistry, Zhejiang Sci-Tech University Hangzhou 310008 P. R. China cuiliang@hnu.edu.cn.
² Biomaterials Research Center, School of Biomedical Engineering, Southern Medical University Guangzhou 510515 P. R. China jhuangaf@connect.ust.hk.

Abstract

An innovative 2'-O-methyl molecular beacon (MB) has been designed and prepared with improved thermal stability and unique nuclease resistance. The employment of 2'-O-methyl MBs helps efficiently suppress the background signal, while DNase I is responsible for the signal amplification and elimination of sticky-end pairing. The coupled use of 2'-O-methyl MBs and DNase I makes it possible to develop an enzyme-aided strategy for amplified detection of DNA targets in a sensitive and specific fashion. The analysis requires only mix-and-measure steps that can be accomplished within half an hour. The detection sensitivity is theoretically determined as 27.4 pM, which is nearly 200-fold better than that of the classic MB-based assay. This proposed sensing system also shows desired selectivity. All these features are of great importance for the design and application of MBs in biological, chemical, and biomedical fields.