Gene Expression on DNA Biochips Patterned with Strand-Displacement Lithography
Gene Expression on DNA Biochips Patterned with Strand-Displacement Lithography
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DOI：
10.1002/anie.201800281
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发表时间：
2018-04-16
影响因子：
16.6
通讯作者：
Simmel, Friedrich C.
中科院分区：
文献类型：
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作者：
Pardatscher, Guenther;Schwarz-Schilling, Matthaeus;Simmel, Friedrich C.
DNA分子的光刻图案化使无细胞遗传电路在良好控制的实验条件下的空间组织成为可能。在这里，我们提出了一种生物相容性，基于DNA的抗蚀剂称为Bephore，这是基于市售的组件，可以通过光和电子束光刻图案。图案化机制是基于紫外线或电子对化学修饰的DNA发夹的切割，以及随后的链置换反应。所有步骤都在水溶液中进行，并且不需要抗蚀剂的化学显影，这使得光刻工艺稳健且生物相容。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。作为一个应用程序，我们展示了区室化，芯片上的基因表达从三个顺序固定的DNA模板，导致三个空间分辨的蛋白质表达梯度。
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.