Collaborative Research: CRISPR-SERS system for rapid and ultrasensitive detection of foodborne bacterial pathogens

合作研究：用于快速、超灵敏检测食源性细菌病原体的 CRISPR-SERS 系统

• 批准号: 2031242
• 负责人: Xue Gao
• 金额: $ 15万
• 依托单位: William Marsh Rice University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2031242&HistoricalAwards=false
• 关键词: Collaborative; Research; CRISPR; SERS; system

Bacterial pathogens and their toxins are the most common causes of foodborne illnesses. Detecting pathogenic bacterial contaminants in complex matrices (e.g., fresh produce) is an essential step to ensure food safety. One of the major challenges for detecting those contaminants lies in the lack of rapid and ultrasensitive detection methods. This multidisciplinary project aims to develop innovative biosensors that can rapidly and accurately screen a large number of samples for pathogenic contaminants. This project is integrated with outreach activities offered through the STEMette Summer Camp and Engineering High School Day to introduce K-12 students to the state-of-the-art biosensor technology. In addition, new course modules based on the results of this project are incorporated into existing undergraduate and graduate courses at both the University of Missouri and Rice University. This project includes three research objectives: (1) design a clustered, regularly interspaced, short palindromic repeats-associated system (CRISPR-Cas) to detect foodborne bacterial pathogens, (2) develop an ultrasensitive SERS system in a microfluidic device, and (3) integrate CRISPR with SERS in the microfluidic device for rapid, ultrasensitive, and selective detection of foodborne bacterial pathogens. This research discovers, contributes to, and supplements the fundamental understanding of essential principles for engineering CRISPR-based next-generation biosensors. By coupling CRISPR with SERS, this study also creates a new sensing mechanism that has the potential to achieve detection with single-copy sensitivity and single-base selectivity.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

细菌病原体及其毒素是食源性疾病的最常见原因。检测复杂基质（例如新鲜农产品）中的致病细菌污染物是确保食品安全的重要步骤。检测这些污染物的主要挑战之一在于缺乏快速和超灵敏的检测方法。这个多学科项目旨在开发创新的生物传感器，能够快速、准确地筛选大量样品中的致病污染物。 该项目与 STEMette 夏令营和工程高中日提供的外展活动相结合，向 K-12 学生介绍最先进的生物传感器技术。此外，基于该项目结果的新课程模块已纳入密苏里大学和莱斯大学现有的本科生和研究生课程。该项目包括三个研究目标：(1) 设计一个簇状、规则间隔、短回文重复序列相关系统 (CRISPR-Cas) 来检测食源性细菌病原体，(2) 在微流体装置中开发超灵敏的 SERS 系统，以及 (3) ）将 CRISPR 与 SERS 集成到微流体装置中，以快速、超灵敏和选择性地检测食源性细菌病原体。这项研究发现、贡献并补充了对基于 CRISPR 的下一代生物传感器工程基本原理的基本理解。通过将 CRISPR 与 SERS 结合，这项研究还创建了一种新的传感机制，有可能实现单拷贝灵敏度和单碱基选择性的检测。该奖项反映了 NSF 的法定使命，并通过使用基金会的评估，认为值得支持。智力价值和更广泛的影响审查标准。

项目成果

CRISPR-Cas12a Biosensor Array for Ultrasensitive Detection of Unamplified DNA with Single-Nucleotide Polymorphic Discrimination

CRISPR-Cas12a 生物传感器阵列，用于通过单核苷酸多态性辨别超灵敏检测未扩增的 DNA

• DOI: 10.1021/acssensors.2c02495
• 发表时间: 2023-04
• 期刊: ACS Sensors
• 影响因子: 8.9
• 作者: Weng, Zhengyan;You, Zheng;Li, Huijie;Wu, Guangfu;Song, Yang;Sun, He;Fradlin, Ava;Neal;Lin, Mengshi;Gao, Xue;et al
• 通讯作者: et al

Engineered LwaCas13a with enhanced collateral activity for nucleic acid detection

工程化 LwaCas13a，具有增强的核酸检测附带活性

• DOI: 10.1038/s41589-022-01135-y
• 发表时间: 2022-09-22
• 期刊: Nature Chemical Biology
• 影响因子: 14.8
• 作者: Jie Yang;Yang Song;Xiangyu Deng;Jeffrey A Vanegas;Zheng You;Yuxuan Zhang;Zhengyan Weng;L. Avery;K. Dieckhaus;Advaith Peddi;Yang Gao;Yi Zhang;Xue Gao
• 通讯作者: Xue Gao

CRISPR‐Cas Biochemistry and CRISPR‐Based Molecular Diagnostics

CRISPR-Cas 生物化学和基于 CRISPR-的分子诊断

• DOI: 10.1002/anie.202214987
• 发表时间: 2023-01
• 期刊: Angewandte Chemie International edition in English
• 作者: Weng, Zhengyan;You, Zheng;Yang, Jie;Mohammad, Noor;Lin, Mengshi;Wei, Qingshan;Gao, Xue;Zhang, Yi
• 通讯作者: Zhang, Yi