DNA binding and reactivity assays based on in-frame protein expression

Abstract

We report herein the development of a DNA binding and reactivity assay methodology based on the in-frame protein expression scheme. The stringent requirement for the precise in vivo translation of sequence information into an amplified readout signal, coupled with a strand displacement-based sequence recognition strategy and antibiotic-conferred channel selection capability, has allowed for the elaboration of a DNA diagnostic protocol with high sensitivity, high selectivity, and multiple-target detection capacity. The efficacy of the proposed DNA detection scheme is further demonstrated through the assay on a genomic target. Importantly, the uniquely structured signal blocking/generation DNA architecture provides a generic handle for the examination of DNA reaction, DNA-small molecule binding, and DNA-protein binding events. The exceptional versatility associated with the one-platform system is anticipated to contribute to the screening and therefore understanding of a diversity of DNA-centered biological phenomena.