Designer virus-inspired proteins drive the manufacturing of more effective and safer gene-delivery systems as well as simpler models to study viral assembly. However, the self-assembly of engineered viromimetic proteins on specific nucleic acid templates, a distinctive viral property, has proved difficult. Inspired by viral packaging signals, we harness the programmability of CRISPR-Cas12a to direct the nucleation and growth of a self-assembling synthetic polypeptide into virus-like particles (VLP) on specific DNA molecules. Positioning up to ten nuclease-dead Cas12a (dCas12a) proteins along a 48.5 kbp DNA template triggers particle growth and full DNA encapsidation at limiting polypeptide concentrations. Particle growth rate was further increased when dCas12a was dimerized with a polymerization silk-like domain. Such improved self-assembly efficiency allows for discrimination between cognate versus non-cognate DNA templates by the synthetic polypeptide. Our CRISPR-guided VLPs could help develop programmable bio-inspired nanomaterials with applications in biotechnology as well as viromimetic scaffolds to improve our understanding of viral self-assembly.

中文翻译：

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CRISPR引导的人工病毒样核衣壳可编程自组装

受设计者病毒启发的蛋白质推动制造更有效，更安全的基因传递系统以及研究病毒装配的更简单模型。然而，已证明在特定核酸模板上工程改造的拟病毒蛋白的自组装，独特的病毒性质是困难的。受病毒包装信号的启发，我们利用CRISPR-Cas12a的可编程性将自组装合成多肽的成核和生长定向到特定DNA分子上的病毒样颗粒（VLP）中。沿着48.5 kbp DNA模板放置多达十个核酸酶死的Cas12a（dCas12a）蛋白会在有限的多肽浓度下触发颗粒生长和完整的DNA衣壳化。当dCas12a用聚合丝状结构域二聚时，颗粒生长速率进一步提高。这种提高的自组装效率允许通过合成多肽区分同源DNA模板和非同源DNA模板。我们的CRISPR引导VLP可以帮助开发可编程的，受生物启发的纳米材料，并将其应用于生物技术以及拟病毒支架中，以增进我们对病毒自组装的了解。