Overview
In order to bring our biological system to reality, we followed the Design–Build–Test–Learn (DBTL) cycle. By applying this engineering approach through multiple iterations, we not only implemented our system successfully but also significantly optimized its performance, both in silico and in the laboratory.
To view different cycles, first click on any of the Design, Build, Test, or Learn sections. Once a section is open, use the arrows that appear to move forward or backward between the cycles. There are a total of 4 Wet Lab DBTL cycles and an additional Dry Lab cycle.
4th engineering cycle
Based on what we learned from the first cycle, we redesigned hairpin probe H0 to achieve higher thermodynamic stability while retaining a poly-uracil loop as the collateral cleavage site for Cas13a. The goal is the initiation of CHA only in the presence of biomarkers.
1st Engineering Cycle
Before validating our full system, we first wanted to assess whether the chosen hairpin sequences would be effective for the Catalytic Hairpin Assembly (CHA) reaction.