Using allosteric transcription factor gated Cas12a activation for metal ion and small molecule biomarker detection
OA Version
Citation
Abstract
Biosensors are pivotal in addressing global challenges in healthcare, environmental monitoring, and diagnostics by providing rapid, sensitive, and portable detection solutions. This thesis explores the integration of allosteric transcription factors (aTFs) and Cas12a to create a novel biosensing platform, leveraging the ligand specificity of aTFs and the robust signal amplification of Cas12a’s trans-cleavage activity. The platform focuses on detecting biomarkers such as N-Acetylneuraminic acid (Neu5Ac) and heavy metals like lead, cadmium, and copper, using corresponding aTFs—NanR, CadC, and CsoR. Experimental evaluations address heavy metal interference on Cas12a activity, aTF gating efficiency, and variability among Cas12a orthologs. Our findings showed differences in heavy metal interference on different Cas12a orthologs and the ability for aTFs to gate Cas12a activation. Future development and expansion of the platform could lead to innovative solutions for precise diagnostics and environmental monitoring, addressing challenges in resource-limited and field-deployable settings.
Description
2025