Synthetic Gene Circuit-Based Assay with Multilevel Switch Enables Background-Free and Absolute Quantification of Circulating Tumor DNA
作者机构：The Key Laboratory of Biomedical Information Engineering of Ministry of EducationSchool of Life Science and TechnologyXi’an Jiaotong UniversityXi’an 710049P.R.China Bioinspired Engineering and Biomechanics Center(BEBC)Xi'an Jiaotong UniversityXi’an 710049P.R.China TFX GroupXi'an Jiaotong University Institute of Life HealthXi'an 710049P.R.China Department of Mechanical EngineeringKyung Hee UniversityYongin 17104Republic of Korea Department of Transfusion MedicineSouthwest HospitalThird Military Medical University(Army Medical University)Chongqing 400038P.R.China Department of Medical OncologyThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'an 710061P.R.China
出 版 物：《Research》 (研究（英文）)
年 卷 期：2023年第2023卷第4期
页 面：821-832页
核心收录：
学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学]
基 金：This work was supported by the National Natural Science Foundation of China(21904104) the Shaanxi Sanqin Scholars Innovation Team Support Plan(xtr062021001) the General Financial Grant from the China Postdoctoral Science Foundation(2020M673418)
主 题：Circulating specificity absolute
摘 要：Circulating tumor DNA(ctDNA)detection has found widespread applications in tumor diagnostics and treatment,where the key is to obtain accurate quantification of ***,this remains challenging due to the issue of background noise associated with existing *** this work,we developed a synthetic gene circuit-based assay with multilevel switch(termed CATCH)for background-free and absolute quantification of *** multilevel switch combining a small transcription activating RNA and a toehold switch was designed to simultaneously regulate transcription and translation processes in gene circuit to eliminate background ***,such a multilevel switch-based gene circuit was integrated with a Cas9 nickase H840A(Cas9n)recognizer and a molecular beacon reporter to form CATCH for ctDNA *** CATCH can be implemented in one-pot reaction at 35°C with virtually no background noise,and achieve robust absolute quantification of ctDNA when integrated with a digital chip(i.e.,digital CATCH).Finally,we validated the clinical capability of CATCH by detecting drug-resistant ctDNA mutations from the plasma of 76 non-small cell lung cancer(NSCLC)patients,showing satisfying clinical sensitivity and *** envision that the simple and robust CATCH would be a powerful tool for next-generation ctDNA detection.
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