Development of highly sensitive one-pot ERA-CRISPR assays for on-site detection of CaMV35S promoter and NOS terminator in genetically modified crops
文献类型： 外文期刊
作者： Liang, Qianqian;Wang, Xiaohui;Xie, Qingqing;Cha, Shaozhen;Lei, Jiawen;Cheng, Guojun;Yu, Guanghui;Wu, Yuhua;Zhang, Li
作者机构：
关键词： Genetically modified organisms;CRISPR/Cas12a;Enzymatic recombinase amplification;One-pot;On-site detection
期刊名称： FOOD CONTROL
ISSN： 0956-7135
年卷期： 2025 年 170 卷
页码：
收录情况： SCIE（2025版）
摘要： The integration of nucleic acid isothermal amplification with CRISPR/Cas12a detection technologies has significantly advanced molecular diagnostics. However, the challenge of integrating both assays into a one-pot reaction to reduce operational complexity and the risk of aerosol contamination has persisted. In this study, we developed a one-pot assay that combines these two powerful tools to improve the detection of genetically modified organisms (GMOs). Targeting the widely used Cauliflower Mosaic Virus 35S promoter (P-CaMV 35S) and the nopaline synthase terminator (T-NOS) from Agrobacterium tumefaciens in genetically modified (GM) crops, we employed a definitive screening design (DSD) approach to optimize the balance between ERA amplification and Cas12a activity. This optimization was achieved by enhancing ERA amplification and precisely adjusting the concentrations of Cas12a and other reaction components, resulting in an efficient and streamlined process. The optimized one-pot ERA-CRISPR/Cas12a system achieved a detection sensitivity of 10 copies per reaction for both P-CaMV 35S and T-NOS within 40 min at 40 degrees C, and was capable of detecting GMO content as low as 0.1% in spiked samples. Moreover, with minimal equipment requirements, such as an LED blue light and a smartphone for result interpretation, this method is highly user-friendly. Combining rapid detection, high specificity, and operational simplicity, this system represents a significant advancement in supporting GMO regulation and global trade, and serves as a promising model for the development of future nucleic acid-based assays.
分类号：
- 相关文献
作者其他论文 更多>>
-
Canolol Alleviates Ethanol-Induced Gastric Ulcer by Inhibiting p38 MAPK/NF-κB/NLRP3 Pathway
作者：Ma, Congcong;Zhang, Li;Huang, Qingde;Deng, Qianchun;Huang, Fenghong;Xu, Jiqu
关键词：Canolol;Gastric ulcer;Gastric mucosal defensesystems;Inflammation;Oxidative stress;Apoptosis;p38 MAPK/NF-kappa B/NLRP3 pathway
-
Discovery of Novel Inhibitors Targeting Fungal Chitin Deacetylase via Virtual Screening for Plant Disease Control
作者：Xu, Huan;Li, Yingchen;Cao, Xiaoyan;He, Qi;Jin, Binyan;Dai, Lingjie;Zhang, Xinyuan;Zhang, Xiaoming;Bian, Qiang;Yang, Qing;Zhang, Li
关键词：（2-0-3）plant fungal disease control;chitin deacetylase;pharmacophore;virtual screening
-
Phenethyl isothiocyanate ameliorates liver injuries secondary to inflammatory bowel disease
作者：Ma, Congcong;Zhang, Li;Huang, Qingde;Deng, Qianchun;Huang, Fenghong;Xu, Jiqu
关键词：Phenethyl isothiocyanate; Inflammatory bowel disease; Secondary liver injury; Inflammation; Pyroptosis; Oxidative stress; Heat shock response
-
Complex Landscape Rice Extraction Using Integrated Sentinel-2 Spectral-Temporal-Spatial Imagery and a Hybrid Deep Learning Architecture
作者：Liu, Tianjiao;Duan, Si-Bo;Liu, Niantang;Zhang, Youzhi;Chen, Jiankui;Zhang, Li;Li, Dong
关键词：Feature extraction;Crops;Accuracy;Remote sensing;Transformers;Data mining;Training;Long short term memory;Data models;Vegetation mapping;Active learning;hybrid deep learning model;rice extraction;Sentinel-2 spectral-temporal-spatial imagery
-
Mitigating Cannabidiol's Non-Selective Cytotoxicity via Subcellular Organelle Targeting: Exploring Mitochondrial Targeting Potential
作者：Liu, Genglian;Li, Ru;Gao, Jingwei;Lin, Cong;Li, Hongyuan;Peng, Yinghua;Wang, Hongshuang;Wang, Xiaohui
关键词：
-
Fusarium spp. Associated With Leaf Blight of Hosta (Hosta plantaginea L.) in China
作者：Jiao, Chenyang;Zhang, Li;Xu, Xi;Yang, Xilang;Guo, Lifeng;Wang, Xiangjing;Xiang, Wensheng;Zhao, Junwei
关键词：Fusarium spp.;Hosta;pathogenicity test;phylogenetic analysis
-
Newly isolated bacteria increase the survival of honeybee larvae infected with Paenibacillus larvae
作者：Zhao, Hongyang;Han, Bo;Yang, Sa;Zhao, Danyang;Zhang, Li;Dai, Pingli
关键词：Apis mellifera;American foulbrood;Leuconostoc citreum;Latilactobacillus sakei;antimicrobial peptides