biostudies-literatureCalcines-Cruz CDirecci?n General de Asuntos del Personal Acad?mico, Universidad Nacional Aut?noma de M?xicoUniversity of Texas at AustinWelch FoundationConsejo Nacional de Ciencia y Tecnolog?aNational Institute of General Medical SciencesNIGMS NIH HHS2752-2757https://www.ebi.ac.uk/biostudies/studies/S-EPMC9724498biostudies-literatureUnknown21(7)Designer virus-inspired proteins drive the manufacturing of more effective, safer gene-delivery systems and simpler models to study viral assembly. However, 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 is further increased when dCas12a is dimerized with a polymerization silk-likeNano lettersCRISPR-Guided Programmable Self-Assembly of Artificial Virus-Like Nucleocapsids.PMC9724498R01 GM124141IA200119GM124141F-1016Calcines-Cruz CFinkelstein IJHernandez-Garcia AfalseCRISPR-Guided Programmable Self-Assembly of Artificial Virus-Like Nucleocapsids.Designer virus-inspired proteins drive the manufacturing of more effective, safer gene-delivery systems and simpler models to study viral assembly. However, 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 is further increased when dCas12a is dimerized with a polymerization silk-like2021-01-01T00:00:00Z2021 Apr2025-04-19T05:54:46.14Z2025-04-19T05:54:46.14ZS-EPMC97244983372981310.1021/acs.nanolett.0c04640