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bioRxiv · 10.1101/2020.10.17.343996

CRISPR-guided programmable self-assembly of artificial virus-like nucleocapsids

Calcines Cruz, C.·FInkelstein, I. J.·Hernandez Garcia, A.
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Abstract

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.

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Calcines Cruz, C., FInkelstein, I. J., Hernandez Garcia, A.. 2020-10-17. CRISPR-guided programmable self-assembly of artificial virus-like nucleocapsids. https://doi.org/10.1101/2020.10.17.343996

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