<b>Reconfiguring the split guide/target active TtAgo for non-amplified multiplex detection of tumor nucleic acids and protein biomarkers</b>

In this strategy, otherwise catalytically inert nucleic acid fragments are reconstituted into functional TtAgo components only in the presence of the target.

Consequently, the target directly participates in forming the active guide or target structure, transducing target recognition into programmable TtAgo cleavage signals without the need for nucleic acid amplification. Furthermore, we introduced FEN1-mediated molecular cleavage to accommodate heterogeneous DNA/RNA termini, and extended the split-guide concept to protein-bound enzymatic biomarkers by designing guide sequences containing apurinic/apyrimidinic (AP) sites that are activated upon APE1-mediated cleavage.</b></p>