Recipient Organization
The Regents of University of California
200 University Office Building
Riverside,CA 92521
Performing Department
(N/A)
Non Technical Summary
California's avocado industry, a $400 million cornerstone of the state's agricultural sector, faces a significant challenge - Avocado Sunblotch Viroid disease (ASBVd). This insidious pathogensilently infect avocado trees, often without outward symptoms. The economic consequences are devastating, with infected trees experiencing substantial yield reduction. Current methods for detecting ASBVd are impeded by their complexity and time-consuming nature, rendering them impractical for large-scale monitoring efforts. This proposal seeks to develop a transformative technology that leverages a unique combination of cutting-edge tools, including ultra-sensitive nanoprobes, the revolutionary CRISPR gene-editing technique, and high-throughput lab-on-a-chip devices. This innovative system has the remarkable potential to simultaneously analyze hundreds of avocado samples within minutes, detecting ASBVd at incredibly low concentrations. The benefits of this innovative technologyextend far beyond the safeguarding of California's avocado industry. Ithas the potential to serve as a model for the development of rapid, reliable, and cost-effective detection methods for a wide range of plant and animal diseases, contributing significantly to the advancement of agricultural disease management practices.
Animal Health Component
(N/A)
Research Effort Categories
Basic
60%
Applied
(N/A)
Developmental
40%
Goals / Objectives
This research proposes the development of a novel High-throughput CRISPR-assisted Nanostress Sensing (HCANS) system for the efficient and large-scale surveillance of Avocado Sunblotch Viroid Disease (ASBVd) in California's avocado production sector. This integrated platform leverages cutting-edge technologies: (i) ultra-sensitive and low-interference Surface-Enhanced Raman Spectroscopy (SERS) nanostress sensing, (ii) rapid and highly-specific CRISPR technology for targeted viroid identification, and (iii) high-throughput microfluidics for automated sample processing and analysis. This innovative system will enable the rapid and simultaneous detection of ASBVd across multiple tree samples within minutes, significantly enhancing current surveillance capabilities. The long-term vision is to develop a fully-automated and intelligent HCANS platform capable of multiplexed viral disease detection across various plant and animal species. The successful development of HCANS will not only mitigate the substantial threat ASBVd poses to the California avocado industry but also contribute to the broader protection of the nation's agricultural and food supply sectors.
Project Methods
This proposal introduces a novel High-throughput CRISPR-assisted Nanostress Sensing (HCANS) system for rapid, reliable, and cost-effective detection of Avocado Sunblotch Viroid Disease (ASBVd) in California's avocado production sector. The HCANS system leverages SERS nanostress sensing, a noninvasive technique that correlates peak shifts in a Raman reporter molecule with its mechanical deformation caused by stressor molecule binding/unbinding. This method offers advantages like high sensitivity, minimal background noise, and label-free operation. High-performance silver nanocrystal (AgNC) substrates will be employed to significantly enhance the signal of the Raman reporter. These AgNCs exhibit broad absorption spectra, allowing excitation with a near-infrared laser to minimize autofluorescence interference. Additionally, their cost-effective and scalable synthesis facilitates precise control over their size, shape, and surface chemistry for optimal performance in large-scale disease screening applications. The HCANS system achieves high detection specificity through the programmable nature of the CRISPR-Cas system. Specific binding with target DNA/RNA triggers the Cas complex to cleave a DNA molecule attached to the Raman reporter. This process restores the reporter's bonds, inducing characteristic peak shifts in the Raman spectrum that correlates with the viroid concentration in the sample. Finally, the HCANS system is integrated with microfluidics chips for high-throughput, cost-effective, and simplified one-step detection. This approach enables parallel analysis of hundreds of samples while minimizing reagent consumption.