Manufacturing Reusable NAND Logic Gates and Their Initial Circuits for DNA Nanoprocessors

Abstract DNA‐based computers can potentially analyze complex sets of biological markers, thereby advancing diagnostics and the treatment of diseases. Despite extensive efforts, DNA processors have not yet been developed due, in part, to limitations in the ability to integrate available logic gates into circuits. We have designed a NAND gate, which is one of the functionally complete set of logic connectives. The gate's design avoids stem‐loop‐folded DNA fragments, and is capable of reusable operations in RNase H‐containing buffer. The output of the gate can be translated into RNA‐cleaving activity or a fluorescent signal produced either by a deoxyribozyme or a molecular beacon probe. Furthermore, three NAND‐gate‐forming DNA strands were crosslinked by click chemistry and purified in a simple procedure that allowed ≈10 13 gates to be manufactured in 16 h, with a hands‐on time of about 30 min. Two NAND gates can be joined into one association that performs a new logic function simply by adding a DNA linker strand. Approaches developed in this work could contribute to the development of biocompatible DNA logic circuits for biotechnological and medical applications.