One treatment of HIV/AIDS is antiretroviral therapy (ART). Antiretroviral therapy helps people with HIV live longer, healthier lives and reduces the risk of HIV transmission. Clinical trial data have shown that once-a-month long-acting injectables suppress HIV comparably to the standard daily oral regimens. To accurately monitor the therapeutic progress and to assure patients’ confidence in the long-period treatment, a technology that is capable of testing certain biomarkers for HIV is essential. This CAREER proposal seeks to study the fundamental properties of the CRISPR technology for applications in ultrasensitive molecular diagnostics and to design and develop a chip-based, smartphone-readable CRISPR biosensor for monitoring the therapeutic progress of HIV treatments. The success of the project will significantly improve the quality and outcome of long-acting HIV treatment. By integrating research and education, this project will increase a diverse STEM workforce by attracting, retaining, and training students in the interdisciplinary field of engineering and biotechnology.
The research objective of this CAREER proposal is to study the unique characteristics of CRISPR trans-nuclease and convert the genome-editing CRISPR-Cas platform into next-generation, rapid, and ultrasensitive biosensors. In pursuit of this goal, the enzymatic activity and kinetics of CRISPR Cas proteins will be systematically studied, and a new kinetic model for the one-pot CRISPR diagnostic reaction will be proposed. This project will also explore the transformation of the solution-based CRISPR assay into a chip-based biosensor that can be optically detected by a smartphone fluorescence microscope to ensure high detection sensitivity to the sub-attomolar level. This new sensing mechanism on a chip will eliminate the need for pre-amplifications of the samples to significantly simplify and shorten the steps of current CRISPR diagnostics. The smartphone-integrated HIV testing platform will allow for both viral load measurement and computational predication, through the incorporation of a viral dynamic model into a custom-developed smartphone application interface. The project will innovate the current toolbox for POC HIV viral load testing, which is one of the major roadblocks in halting the HIV epidemic and enhancing treatment. The results of this project will provide new insights into the detailed mechanism of trans-cleavage of CRISPR Cas proteins. The successful development of such distributed and connected POC sensors will be very timely in the new wave of long-acting injectable and implantable HIV drugs. We expect the developed sensory methodology can also be broadly applied to the detection of many other diseases in resource-limited settings in the future.
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.
