Enhancing 3D DNA Walker-Induced CRISPR/Cas12a Technology for Highly Sensitive Detection of ExomicroRNA Associated with Osteoporosis

作者全名："Luo, Lijuan; Dong, Fang; Li, Dandan; Li, Xinmin; Li, Xinyu; Fan, Yunpeng; Qi, Caihong; Luo, Jinyong; Li, Li; Shen, Bo"

作者地址："[Luo, Lijuan] Chongqing Med Univ, Affiliated Hosp 3, Dept Lab Med, Chongqing 401120, Peoples R China; [Dong, Fang] Chongqing Med Univ, Affiliated Hosp 1, Dept Gerontol, Branch 1, Chongqing 400000, Peoples R China; [Li, Dandan] Chongqing Med Univ, Affiliated Hosp 2, Dept Lab Med, Chongqing 400072, Peoples R China; [Li, Xinmin; Fan, Yunpeng; Shen, Bo] Chongqing Hosp Tradit Chinese Med, Chongqing Key Lab Sichuan Chongqing Coconstruct Di, Chongqing 400021, Peoples R China; [Li, Xinyu; Fan, Yunpeng; Luo, Jinyong] Chongqing Med Univ, Minist Educ, Coll Lab Med, Key Lab Clin Lab Diagnost, Chongqing 400016, Peoples R China; [Qi, Caihong; Li, Li] Chongqing Gen Hosp, Dept Lab Med, Chongqing 401147, Peoples R China"

通信作者："Shen, B (通讯作者)，Chongqing Hosp Tradit Chinese Med, Chongqing Key Lab Sichuan Chongqing Coconstruct Di, Chongqing 400021, Peoples R China.; Li, L (通讯作者)，Chongqing Gen Hosp, Dept Lab Med, Chongqing 401147, Peoples R China."

来源：ACS SENSORS

ESI学科分类：CHEMISTRY

WOS号：WOS:001181188800001

JCR分区：Q1

影响因子：8.2

年份：2024

卷号：9

期号：3

开始页：1438

结束页：1446

文献类型：Article

关键词：fluorescent biosensor; CRISPR/Cas12atechnology; 3D DNA walker; exomiRNA analysis; osteoporosis

摘要："Exosomal microRNAs (exomiRNAs) have emerged as promising biomarkers for the early clinical diagnosis of osteoporosis. However, their limited abundance and short length in peripheral blood present significant challenges for the accurate detection of exomiRNAs. Herein, we have designed and implemented an efficacious fluorescence-based biosensor for the highly sensitive detection of exomiRNA associated with osteoporosis, leveraging the enhancing 3D DNA walker-induced CRISPR/Cas12a technology. The engineered DNA walker is capable of efficiently transforming target exomiRNA into amplifying DNA strands, thereby enhancing the sensitivity of the developed biosensor. Concurrently, the liberated DNA strands serve as activators to trigger Cas12a trans-cleavage activity, culminating in a significantly amplified fluorescent signal for the highly sensitive detection of exomiRNA-214. Under optimal conditions, the devised technology demonstrated the capacity to detect target exomiRNA-214 at concentrations as low as 20.42 fM, encompassing a wide linear range extending from 50.0 fM to 10.0 nM. Moreover, the fluorescence-based biosensor could accurately differentiate between healthy individuals and osteoporosis patients via the detection of exomiRNA-214, which was in agreement with RT-qPCR results. As such, this biosensing technology offers promise as a valuable tool for the early diagnosis of osteoporosis."

基金机构："Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University [82002257]; National Natural Science Foundation of China [cstc2021jcyj-msxmX0325]; Natural Science Foundation Project of Chongqing [2022MSXM106, 2023QNXM028, 2023MSXM035]; Chongqing Medical Scientific Research Project-Joint project of Chongqing Health Commission and Science and Technology Bureau [2023MD734120]; China Postdoctoral Science Foundation [kryc-yq-2117]; Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University"

基金资助正文："This study was supported by the National Natural Science Foundation of China (82002257), the Natural Science Foundation Project of Chongqing (cstc2021jcyj-msxmX0325), Chongqing Medical Scientific Research Project-Joint project of Chongqing Health Commission and Science and Technology Bureau (2022MSXM106, 2023QNXM028, and 2023MSXM035), the China Postdoctoral Science Foundation (Certificate Number: 2023MD734120), and Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University (kryc-yq-2117)."