Dynamic Process in SARS-CoV-2 Replication-Transcription Using Single-Molecule Magnetic Tweezers Technology
10.16156/j.1004-7220.2025.04.014
- VernacularTitle:基于单分子磁镊技术研究SARS-CoV-2复制转录的动态过程
- Author:
Qianhong GUO
1
;
Xiaomin CHEN
;
Zhiqin ZHENG
;
Jun FAN
Author Information
1. 西南科技大学生命科学与工程学院,四川绵阳 621010
- Publication Type:Journal Article
- Keywords:
single-molecule magnetic tweezers;
worm-like chain model;
RNA-dependent RNA polymerase;
replicative transcription
- From:
Journal of Medical Biomechanics
2025;40(4):895-901
- CountryChina
- Language:Chinese
-
Abstract:
Objective To elucidate the kinetic characteristics of viral replication and transcription,an in vitro model of viral replication and transcription was established.Utilizing single-molecule magnetic tweezers technology,the dynamic process of SARS-CoV-2 RNA-dependent RNA polymerase(RdRp)during in vitro replication and transcription was investigated.Methods The force field of the single-molecule magnetic tweezer system was corrected using DNA fragments,followed by the construction of RNA fragments to explore the kinetics of RdRp replication and transcription in vitro.Results The force field calibration results were consistent with the worm-like chain model(WLC).The ssRNA strand was found to be approximately 0.3 pm longer than the dsRNA strand and could be stably extended under a 30 pN force field.The average synthesis rate of RdRp extension was determined to be 3.27 nt/s,with an average processivity of 886 nt.Conclusions By implementing force calibration in single-molecule magnetic tweezers,the real-time tracking of RdRp kinetics during the full-cycle replication-transcription process(initiation,elongation,and termination)in vitro was achieved,thereby constructing a mechanistic model of RdRp-driven nucleic acid synthesis.This study provides a basis for further investigating the kinetics of viral RdRp in physiological processes including replication,transcription,and backtracking under varied in vitro environments using single-molecule magnetic tweezers,and establishes a single-molecule manipulation framework for evaluating the effects of therapeutic compounds on viral replication-transcription processes in vitro.
