Enzyme-driven converter and amplifier for inert double-stranded DNA without sequence restrictions

DNA molecules in life play a crucial role in various fields. However, DNA-based technologies confront significant difficulties in dealing with inert double-stranded DNA (dsDNA) with a blunt end without specific sequences. In this work, we utilized Thermus thermophilus Argonaute (TtAgo) and Vent DNA polymerase to construct an enzyme-driven device to convert and amplify arbitrary blunt-ended dsDNA into single-stranded DNA (ssDNA), thereby adapting to subsequent functional units. Based on the excellent performance of the dsDNA to ssDNA converter and amplifier (DSCA), inert dsDNA could be effectively identified. Additionally, DSCA accomplishes information interception and transmission, addressing the problem of dsDNA input response in DNA circuits and clinical diagnosis. Given these facts, this DSCA is expected to serve as a broad toolbox for the information transformation and amplification of inert dsDNA, prospering DNA circuit operations and biosensing