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

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.