journal article·2026-03Open access

Integrated Electroporated‐Lysis Electrochemical Platform Enables Sensitive and Rapid EV Protein and miRNA Profiling Based on Multiplex‐Responsive CRISPR/Cas12a

Rui Fan, Yihang Tong, Shihua Luo, Yangfen He, Chao Yang, Wenbin Li, Jieyan Liu, Jiezhen Pan, Yiping Zhu, Xiaohe Zhang, Junfang Zhu, Yitong Zhu, Yuhang Guo, Ling Li, Bo Situ, Xiaohui Yan, Wen Ma, Lingqian Chang, Ye Zhang

Small· Volume 22· Issue 13

Abstract

ABSTRACT Proteins and miRNAs in extracellular vesicles (EVs) have emerged as crucial biomarkers for tumor diagnosis. While CRISPR/Cas12a‐based platforms have shown great promise in nucleic acid and protein detection, their susceptibility to off‐target activation and structural instability remains a significant limitation. Here, we have developed an electroporation‐lysis electrochemical platform integrated with DNA cube‐cage‐locked CRISPR/Cas12a (DC‐Cas12a), termed EL‐DC‐Cas12a. This platform utilizes an electric field to rapidly lyse EVs, releasing their internal proteins and miRNAs. These released molecules then activate the DC‐Cas12a system, thereby triggering the displacement of two distinct crRNA/Cas12a complexes that correspond to EV proteins and miRNAs, respectively. These complexes then specifically recognize and cleave electrochemical probes, generating quantifiable electrochemical signals that enable synchronous and accurate analysis of the two biomarkers. The integrated workflow for EV lysis and detection can be completed within 40 min, greatly simplifying the overall operation. The detection limits (LOD) of this platform for EV PD‐L1 protein and miR‐1246 were 5.44 × 10 4 particles/mL and 3.59 × 10 3 particles/mL, respectively. Moreover, by applying machine learning algorithms to analyze the EV‐associated proteins and miRNAs profiling, the platform demonstrated a diagnostic accuracy of 98.3% in distinguishing healthy donors from early‐stage GC patients, and 99% in differentiating early‐stage from advanced‐stage GC patients in a clinical gastric cancer cohort. Therefore, the proposed platform offers a promising strategy for multiplexed detection of EV biomarkers and precise discrimination of GC.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: Extracellular vesicles in disease · Advanced biosensing and bioanalysis techniques · CRISPR and Genetic Engineering

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Department of Laboratory Medicine Guangdong Provincial Key Laboratory of Precision Medical Diagnostics Guangdong Engineering and Technology Research Center for Rapid Diagnostic Biosensors Guangdong Provincial Key Laboratory of Single‐cell and Extracellular Vesicles Nanfang Hospital Southern Medical University Guangzhou P. R. China

School of Medical Technology Guangdong Medical University Dongguan P. R. China

School of Engineering Medicine Beihang University Beijing P. R. China

Qingdao Research Institute Beihang University Qingdao P. R. China

Center for Clinical Laboratory Diagnosis and Research Affiliated Hospital of Youjiang Medical University for Nationalities Baise Guangxi P. R. China

Key Laboratory of Research on Clinical Molecular Diagnosis for High Incidence Diseases in Western Guangxi of Guangxi Higher Education Institutions Affiliated Hospital of Youjiang Medical University for Nationalities Baise Guangxi P. R. China

The Affiliated Traditional Chinese Medicine Hospital Guangzhou Medical University Guangzhou P. R. China

Medical Research Center of Nanfang Hospital Nanfang Hospital Guangdong Provincial Key Laboratory of Proteomics School of Basic Medical Sciences Southern Medical University Guangzhou P. R. China

Shenzhen Hospital Southern Medical University Shenzhen Guangdong P. R. China

Funding

National Key Research and Development Program of China · National Natural Science Foundation of China · Southern Medical University · China Postdoctoral Science Foundation · National Science Fund for Distinguished Young Scholars

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