The bottom-up engineering of artificial cells requires a reconfigurable cytoskeleton that can organize at distinct locations and dynamically modulate its structural and mechanical properties. Here, inspired by the vast array of actin-binding proteins and their ability to reversibly crosslink or bundle filaments, we have designed a library of peptide–DNA crosslinkers varying in length, valency and geometry. Peptide filaments conjoint through DNA hybridization give rise to tactoid-shaped bundles with tunable aspect ratios and mechanics. When confined in cell-sized water-in-oil droplets, the DNA crosslinker design guides the localization of cytoskeletal structures at the cortex or within the lumen of the synthetic cells. The tunable spatial arrangement regulates the passive diffusion of payloads within the droplets and complementary DNA handles allow for the reversible recruitment and release of payloads on and off the cytoskeleton. Heat-induced reconfiguration of peptide–DNA architectures triggers shape deformations of droplets, regulated by DNA melting temperatures. Altogether, the modular design of peptide–DNA architectures is a powerful strategy towards the bottom-up assembly of synthetic cells.

人造细胞的自下而上工程需要可重构的细胞骨架，可以在不同的位置组织并动态调节其结构和机械特性。在这里，受到大量肌动蛋白结合蛋白及其可逆交联或成束细丝的能力的启发，我们设计了一个长度、化合价和几何形状各异的肽-DNA交联剂库。肽丝通过 DNA 杂交结合在一起，形成具有可调节纵横比和力学结构的类晶状纤维束。当被限制在细胞大小的油包水液滴中时，DNA 交联剂设计引导细胞骨架结构在皮质或合成细胞内腔内的定位。可调的空间排列调节液滴内有效负载的被动扩散，互补DNA手柄允许有效负载在细胞骨架上和外的可逆招募和释放。热诱导的肽-DNA 结构重构会触发液滴的形状变形，并受 DNA 熔化温度的调节。总而言之，肽-DNA 架构的模块化设计是实现合成细胞自下而上组装的强大策略。