Deoxyribonucleic acid (DNA) hydrogels, a three-dimensional (3D) network made from DNA chains, have attracted great attention because of its molecular programmability, excellent biocompatibility and wide biomedical applications. Construction of hydrogel incorporating genetic function is still a challenge because of the limitations in available preparation methods. Herein, we develop a polymerase chain reaction (PCR) based strategy to construct gene integrated hydrogel to mimic the biofunction of nucleus zone. DNA primers were chemically modified by methacrylamide, which were used as modular primers in PCR to hybridize with template plasmid DNA, yielding methacrylamide functionalized gene (Acry-gene). Afterwards, Acry-gene was chemically cross-linked and compressed via free radical polymerization of terminal group methacrylamide to form a three-dimensional gene network, namely gene hydrogel. The gene hydrogel retained the genetic function and expressed protein successfully in a cell free protein expression system. This work provides a general approach for the construction of biofunctional gene hydrogel which mimics bioprocesses, showing great potential in biomedicine and biomimetic fields.

中文翻译：

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通过聚合酶链反应制备仿生基因水凝胶以表达无细胞蛋白

由DNA链制成的三维（3D）网络脱氧核糖核酸（DNA）水凝胶因其分子可编程性，出色的生物相容性和广泛的生物医学应用而备受关注。由于现有制备方法的局限性，构建具有遗传功能的水凝胶仍然是一个挑战。在本文中，我们开发了一种基于聚合酶链反应（PCR）的策略，以构建基因整合的水凝胶来模拟核区的生物功能。DNA引物经甲基丙烯酰胺化学修饰，在PCR中用作模块引物，与模板质粒DNA杂交，产生甲基丙烯酰胺功能化基因（Acry基因）。然后，Acry基因经过化学交联并通过端基甲基丙烯酰胺的自由基聚合反应进行压缩，从而形成三维基因网络，即基因水凝胶。该基因水凝胶保留了遗传功能，并在无细胞蛋白质表达系统中成功表达了蛋白质。这项工作为模仿生物过程的生物功能基因水凝胶的构建提供了一种通用方法，在生物医学和仿生领域显示出巨大的潜力。