Physics-Department and ZNN, Technische Universität München, Am Coulombwall 4a, 85748, Garching, Germany.
Department of Chemical and Biological Physics, The Weizmann Institute of Science, Rehovot, 76100, Israel.
Angew Chem Int Ed Engl. 2018 Apr 16;57(17):4783-4786. doi: 10.1002/anie.201800281. Epub 2018 Mar 15.
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.
DNA 分子的光刻图形化技术可在严格控制的实验条件下实现无细胞遗传回路的空间组织。在这里，我们提出了一种基于 DNA 的生物相容性抗蚀剂“Bephore”，它基于市售的成分，可以通过光和电子束光刻进行图案化。图案化机制基于化学修饰的 DNA 发夹在紫外光或电子下的切割，以及随后的链置换反应。所有步骤都在水溶液中进行，不需要抗蚀剂的化学显影，这使得光刻过程具有鲁棒性和生物相容性。Bephore 非常适合多步光刻工艺，能够以微米精度固定不同类型的 DNA 分子。作为一种应用，我们展示了从三个顺序固定的 DNA 模板进行分隔的、芯片上的基因表达，从而产生了三个空间分辨的蛋白质表达梯度。
