A research team led by Prof. WU Tongbo from School of Pharmacy, Tongji Medical College of HUST, has made groundbreaking progress in CRISPR-Cas12a biosensors through innovative topological designs. Their findings, published in Nucleic Acids Research, demonstrated how structural modifications can significantly enhance the system's detection capabilities.

The study systematically investigated how activator topology—spanning linear, planar, and three-dimensional configurations—affects the trans-cleavage activity of CRISPR-Cas12a. This comprehensive approach revealed that carefully designed topological architectures can boost detection sensitivity while improving operational stability, as compared to conventional linear activators.

Prof. WU's team discovered that suppressing Cas12a's cis-cleavage activity through topological optimization paradoxically enhanced its trans-cleavage performance. This counterintuitive finding led to the development of a planar activator-based Cas12a switch that maintains protracted high activity and achieves exceptional sensitivity. When applied to hypochlorous acid detection, the system demonstrated a remarkable detection limit of 88 nM, outperforming existing chemical probe methods.

The research provides detailed structural insights across multiple dimensions:

· Linear designs incorporating bubbles and hairpins revealed specific activity modulation patterns.

· Planar configurations showed significantly stronger activation than their traditional linear counterparts.

· 3D frameworks improved system stability for potential cell imaging applications.

The study exemplified the success of HUST's talent development pipeline. Co-first authors ZHU Zixuan and LI Xiaolong are students on the Qiming Experimental Training Program of the School of Pharmacy's, where undergraduate research projects seamlessly transitions into graduate-level research programs.

The article link：https://academic.oup.com/nar/article/53/8/gkaf311/8117948