Gene Expression on DNA Biochips Patterned with Strand‐Displacement Lithography

Lithographic patterning of DNA molecules enables spatial organization of cell‐free genetic circuits under well‐controlled experimental conditions. Here, we present a biocompatible, DNA‐based resist termed “Bephore”, which is based on commercially available components and can be patterned by both photo‐ and electron‐beam lithography. The patterning mechanism is based on cleavage of a chemically modified DNA hairpin by ultraviolet light or electrons, and a subsequent strand‐displacement reaction. All steps are performed in aqueous solution and do not require chemical development of the resist, which makes the lithographic process robust and biocompatible. Bephore is well suited for multistep lithographic processes, enabling the immobilization of different types of DNA molecules with micrometer precision. As an application, we demonstrate compartmentalized, on‐chip gene expression from three sequentially immobilized DNA templates, leading to three spatially resolved protein‐expression gradients.