Excellence in Research: 2D Heterostructure Materials Based CRISPR Sensors for Detection of Salmonella and its serotypes

食源性疾病是全世界一个主要的公共卫生问题，开发能够快速和准确地检测这些疾病的方法非常重要。基于CRISPR的系统用于检测这些疾病正变得越来越流行，但它们仍然依赖于靶DNA的扩增。为了解决这一问题，基于具有独特电化学性质的二维MXen基异质结构材料，将开发一种新的传感平台。这一新平台可能在不到15分钟内检测到病原体，比现有平台具有更高的灵敏度和特异性。如果成功，这种CRISPR传感器可以快速检测沙门氏菌和其他病原体，而不需要预扩增步骤。该系统还可以容易和快速地扩展，以检测其他新出现的病原体，并可以帮助决策者规划预防措施。该项目还提供了劳动力发展的机会，包括指导研究生和本科生，提供基于CRISPR的生物传感器纳米制造的研讨会，以及招募一组高中生参加iGEM国际比赛。拟议的项目旨在开发一个新的传感平台，使用基于CRISPR的系统和基于2D MXene的异质结构材料来快速检测沙门氏菌。该项目有三个具体目标：目标1：设计针对沙门氏菌及其血清型的特异性gRNA并评价其特异性；目标2：开发一个官能团连接到dCas9酶上，并将其结合到MXen-石墨烯材料的表面末端基团上；目标3：制备两种基于dCas9和Cas12a酶的CRISPR-MXene-石墨烯传感器，验证传感器的有效性，并研究带隙和电容调节对不同类型MXenes传感机制的影响。该项目的智力价值在于促进了对带隙可调的2D异质结构材料中分子相互作用、化学键和电化学响应的理解。根据拟议研究开发的实验方法还可以帮助在更广泛的社区探索类似的科学问题，包括开发用于快速检测其他病原体的通用平台。如果成功，建议的基于CRISPR的MXene-石墨烯系统可以实现对沙门氏菌及其血清型的快速检测，而不需要预扩增步骤。该系统可以很容易地扩展用于检测其他病原体，并可用于远程临床实验室的护理点应用。该项目也有更广泛的影响，包括支持基础技术、劳动力发展和加强HBCU研究基础设施。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Foodborne diseases are a major public health concern all over the world, and it is important to develop methods that can quickly and accurately detect these diseases. The use of CRISPR-based systems is becoming popular for detecting these diseases, but they still rely on the amplification of target DNA. To address this issue, a new sensing platform will be developed based on 2D MXene-based heterostructure materials, which have unique electrochemical properties. This new platform can potentially detect pathogens in less than 15 minutes with superior sensitivity and specificity over existing platforms. If successful, this CRISPR sensor could enable rapid detection of Salmonella and other pathogenic agents without the need for pre-amplification steps. The system can also be easily and rapidly expanded for the detection of other emerging pathogenic agents and can assist policymakers in planning preventive measures. This project also offers opportunities for workforce development, including mentoring graduate and undergraduate students, offering workshops on nanomanufacturing CRISPR-based biosensors, and recruiting a team of high school students for competition at the iGEM international competition.The proposed project aims to develop a new sensing platform for the rapid detection of Salmonella using CRISPR-based systems and 2D MXene-based heterostructure materials. The project has three specific aims: Aim 1: Design specific gRNAs for Salmonella and its serotypes and evaluate their specificity; Aim 2: Develop a functional group to attach to the dCas9 enzyme and bind it to the surface terminal groups of the MXene-graphene materials; Aim 3: Prepare two types of CRISPR-MXene-graphene sensors based on either dCas9 or Cas12a enzymes, validate the sensors, and investigate the effect of bandgap and capacitance tuning on the sensing mechanism for different types of MXenes. The project's intellectual merit lies in advancing understanding of molecular interactions, chemical bonding, and electrochemical response in 2D heterostructure materials with tunable bandgap. The experimental methods developed from the proposed research can also help explore similar scientific problems in a wider range of the community, including the development of versatile platforms for rapid detection of other pathogenic agents. If successful, the proposed CRISPR-based MXene-graphene system could enable rapid detection of Salmonella and its serotypes without the need for pre-amplification steps. The system can be easily expanded for the detection of other pathogenic agents and can be useful for point-of-care applications in remote clinical laboratories. The project also has broader impacts, including enabling foundational technologies, workforce development, and strengthening HBCU research infrastructure.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.