期刊
BIOSENSORS & BIOELECTRONICS
卷 167, 期 -, 页码 -出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2020.112474
关键词
Exosome; RNA; Isothermal gene amplification; Three-way junction; G-quadruplex; Cancer diagnosis
类别
资金
- Center for BioNano Health-Guard - Ministry of Science and ICT of Korea (MSIT) as Global Frontier Project [H-GUARD_2013M3A6B2078950, H-GUARD_2014M3A6B2060507]
- Bio & Medical Technology Development Program of the National Research Foundation of Korea (NRF) - MSIT [NRF-2018M3A9E2022821]
- NRF - MSIT [NRF-2018R1A2A1A05022355]
- KRIBB Research initiative Program
- Global Ph.D. Fellowship [NRF-2019H1A2A1073468]
Exosomal messenger RNA (mRNA) has emerged as a valuable biomarker for liquid biopsy-based disease diagnosis and prognosis due to its stability in body fluids and its biological regulatory function. Here, we report a rapid one-step isothermal gene amplification reaction based on three-way junction (3WJ) formation and the successful detection of urinary exosomal mRNA from tumor-bearing mice. The 3WJ structure can be formed by the association of 3WJ probes (3WJ-template and 3WJ-primer) in the presence of target RNA. After 3WJ structure formation, the 3WJ primer is repeatedly extended and cleaved by a combination of DNA polymerase and nicking endonuclease, producing multiple signal primers. Subsequently, the signal primers promote a specially designed network reaction pathway to produce G-quadruplex probes under isothermal conditions. Finally, G-quadruplex structure produces highly enhanced fluorescence signal upon binding to thioflavin T. This method provides a detection limit of 1.23 pM (24.6 amol) with high selectivity for the target RNA. More importantly, this method can be useful for the sensing of various kinds of mRNA, including breast cancer cellular mRNA, breast cancer exosomal mRNA, and even urinary exosomal mRNA from breast cancer mice. We anticipate that the developed RNA detection assay can be used for various biomedical applications, such as disease diagnosis, prognosis, and treatment monitoring.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据