Cabello-Garcia Javier, Mukherjee Rakesh, Bae Wooli, Stan Guy-Bart V, Ouldridge Thomas E
Department of Bioengineering, Imperial College London, London, UK.
Imperial College Centre for Engineering Biology, Imperial College London, London, UK.
Nat Chem. 2025 Jun 5. doi: 10.1038/s41557-025-01831-x.
Information propagation by sequence-specific, template-catalysed molecular assembly is a key process facilitating life's biochemical complexity, yielding thousands of sequence-defined proteins from only 20 distinct building blocks. However, exploitation of catalytic templating is rare in non-biological contexts, particularly in enzyme-free environments, where even the template-catalysed formation of dimers is challenging. Typically, product inhibition-the tendency of products to bind to templates more strongly than individual monomers-prevents catalytic turnover. Here we present a rationally designed enzyme-free system in which a DNA template catalyses, with weak product inhibition, the production of sequence-specific DNA dimers. We demonstrate selective templating of nine different dimers with high specificity and catalytic turnover, then we show that the products can participate in downstream reactions, and finally that the dimerization can be coupled to covalent bond formation. Most importantly, our mechanism demonstrates a design principle for constructing synthetic molecular templating systems, a first step towards applying this powerful motif in non-biological contexts to construct many complex molecules and materials from a small number of building blocks.
通过序列特异性、模板催化的分子组装进行信息传播是促进生命生化复杂性的关键过程,仅从20种不同的构建模块就能产生数千种序列定义的蛋白质。然而,催化模板化在非生物环境中很少被利用,特别是在无酶环境中,即使是模板催化的二聚体形成也具有挑战性。通常,产物抑制——产物比单个单体更强烈地结合到模板上的倾向——会阻止催化周转。在这里,我们展示了一个经过合理设计的无酶系统,其中DNA模板以较弱的产物抑制作用催化序列特异性DNA二聚体的产生。我们证明了对九种不同二聚体的选择性模板化具有高特异性和催化周转,然后我们表明产物可以参与下游反应,最后二聚化可以与共价键形成偶联。最重要的是,我们的机制展示了构建合成分子模板系统的设计原则,这是在非生物环境中应用这种强大基序以从少量构建模块构建许多复杂分子和材料的第一步。