CRISPR-Powered DNA Computing and Digital Display.

Clustered regularly interspaced short palindromic repeats (CRISPR) technology has unique specificity for recognizing and cleaving target DNA complementary to the CRISPR guide sequence. Here, we report on a CRISPR-powered DNA computing and digital display system in which programmed DNA targets serve as the input and an ON/OFF fluorescence signal indicates a TRUE/FALSE output. This system allows the establishment of a one-to-one relationship between input and output, enabling multilevel DNA logic computing. Applying pre-CRISPR reactions that selectively maintain or inhibit CRISPR reactivity can further improve the computing capability by expanding input size. In particular, we present a paper-based microfluidic chip with freeze-dried CRISPR reaction mixtures that are programmed to digitally display the results of functional operations, including square, cube, and square-root operations. This strategy allows information decoding and displaying as well, which brings potential in next-generation DNA steganography and cryptography. We envision that the intrinsic orthogonality of CRISPR provides a new paradigm for DNA computing and molecular programming.