In Situ Amplified Mutational mRNA Imaging Using a Spatially Confined CRISPR Nanoplatform
Abstract
ABSTRACT Highly sensitive spatial analysis of RNA mutations is essential for understanding cellular heterogeneity and disease mechanisms. Herein, we developed an integrated CRISPR/Cas13a‐based nanoprobe system for rapid detection of RNA in tissue sections ( In tegrated CRISPR/ Cas 13a‐based RNA R apid D etection, InCasRD). Unlike conventional “always‐on” probes that rely on accumulated probe hybridization, InCasRD leverages the trans‐cleavage activity of Cas13a to achieve spatially confined signal amplification and a high signal‐to‐background ratio (SBR). Using InCasRD, we achieved imaging of multiple target RNAs in tumor cells within 0.5 h of incubation, including mRNA ( survivin ), microRNA (miR‐21), and circular RNA (circ1785). Furthermore, the engineered InCasRD system enabled mapping of RNA mutations, such as the EGFR L858R and ovarian tumor domain ( OTUD ) single‐nucleotide variant (SNV, 23439980 G>T), in tumor tissue sections, thereby facilitating clear tumor boundary delineation. Collectively, InCasRD is a powerful, one‐step tool for in situ RNA analysis with potential for diagnosis and precision medicine.
Article Details
Authors (12)
Wen Zhao
School of Materials Science and Engineering, China University of Petroleum (East China), Qingdao, China.
Ziyue Zheng
Department of Chemistry, State Key Laboratory of Porous Materials for Separation and Conversion, Shanghai Key Laboratory of Molecular Catalysis and Innovative Materials, and iChEM
Rui Li
Haojie Xie
School of Pharmaceutical Sciences Key Laboratory of Advanced Drug Preparation Technologies Ministry of Education China Pingyuan Laboratory Tianjian Laboratory of Advanced Biomedical Sciences State Key Laboratory of Antiviral Drugs Zhengzhou University Zhengzhou Henan China
Haiyun Yu
School of Pharmaceutical Sciences Key Laboratory of Advanced Drug Preparation Technologies Ministry of Education China Pingyuan Laboratory Tianjian Laboratory of Advanced Biomedical Sciences State Key Laboratory of Antiviral Drugs Zhengzhou University Zhengzhou Henan China
Yongyuan Zhang
Yonghua Wu
School of Pharmaceutical Sciences Key Laboratory of Advanced Drug Preparation Technologies Ministry of Education China Pingyuan Laboratory Tianjian Laboratory of Advanced Biomedical Sciences State Key Laboratory of Antiviral Drugs Zhengzhou University Zhengzhou Henan China
Yunqing Yang
Zhenzhong Zhang
School of Pharmaceutical Sciences, Zhengzhou University
Hua Gao
State Key Laboratory of Fine Chemicals, Frontier Science Center for Smart Material
Yanan Li
NHC Key Laboratory of Biotechnology for Microbial Drugs
Kaixiang Zhang
School of Pharmaceutical Sciences, State Key Laboratory of Metabolic Dysregulation and Prevention and Treatment of Esophageal Cancer, State Key Laboratory of Antiviral Drugs, Tianjian Laboratory of Advanced Biomedical Sciences, Pingyuan Laboratory