title: ‘Individual Final Project’ weight: 10 description: ‘Automated Optimization of a DNAzyme–Cas12a Amplified Lead Sensor’ Automated Optimization of a DNAzyme–Cas12a Amplified Lead Sensor Abstract Lead contamination in drinking water remains a major public health problem because even low-level chronic exposure can impair neurological development, cardiovascular health, and overall long-term wellbeing. Existing analytical methods such as ICP-MS are highly sensitive, but they usually require centralized laboratory infrastructure, trained personnel, and expensive instrumentation, which limits their accessibility for decentralized or field-based monitoring. The overall goal of this project is to develop a modular environmental biosensing platform that couples a Pb²⁺-responsive DNAzyme with CRISPR-Cas12a signal amplification in order to generate a rapid and amplified fluorescent readout. The central hypothesis is that a DNAzyme-triggered release of a programmable nucleic acid activator can be linked to Cas12a collateral cleavage to improve sensitivity while preserving modularity. To test this idea, the project is structured into three aims: first, computational design and kinetic modeling of the sensing cascade; second, automated experimental optimization using robotic liquid handling; and third, long-term translation into a portable and modular environmental sensing format. The methods include nucleic acid folding analysis, structural plausibility assessment, kinetic simulation, DNA construct design, and future automated wet-lab optimization. Together, this project aims to establish a scalable biosensing framework for environmental monitoring that is adaptable, programmable, and ultimately deployable outside centralized laboratories.