Scholarship 24/07237-0 - Sistema livre de células, Repetições palindrômicas curtas agrupadas e - BV FAPESP
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Development of simple, low-cost, and field-deployable PAM-free CRISPR-based biosensors for multiplexed nucleic acid detection

Grant number: 24/07237-0
Support Opportunities:Scholarships abroad - Research Internship - Doctorate
Start date until: September 01, 2024
End date until: August 24, 2025
Field of knowledge:Physical Sciences and Mathematics - Chemistry - Analytical Chemistry
Principal Investigator:Lauro Tatsuo Kubota
Grantee:Dagwin Wachholz Júnior
Supervisor: Keith Ian Pardee
Host Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Institution abroad: University of Toronto (U of T), Canada  
Associated to the scholarship:21/09706-0 - Development of a CRISPR/Cas-based electrochemical biosensor for T-Cell Leukemia virus (HTLV-1) and Staphylococcus Aureus bacteria detection with point-of-care devices, BP.DR

Abstract

Early and efficient disease diagnosis is essential for prompt and accurate treatment selection, aiming to prevent complications and increase survival rates. New technologies for detecting pathogens are being developed as alternatives to existing methods, which are often time-consuming and complex. One of the most promising approaches is using CRISPR/Cas systems for nucleic acid detection. Renowned for its remarkable precision in genome editing and programmability capability, CRISPR has found a novel application in biosensing, promising highly sensitive and specific detection of nucleic acids and biomarkers associated with diverse diseases. While CRISPR-based biosensing has shown great advancements, some challenges for point-of-care applications still hinder their practical implementation. To date, most biosensors rely on fluorescent outputs, which are not applicable for resource-limited settings. Further developments are compromised to the design of colorimetric and electrochemical platforms, enabling low-cost and field-deployable diagnostics. Another problem CRISPR systems face is dependence on PAM-containing target sequences, which restrict sequence selection diversity. The present proposal aims to use a developed method in Pardee Lab at the University of Toronto to create a simple and affordable CRISPR biosensor with colorimetric outputs for field application in diagnostics settings. Furthermore, we aim to develop colorimetric multiplexed biosensors for clinically relevant biomarkers using CRISPR-based technology and cell-free systems.

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