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A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors
Published on: October 30, 2016
CRISPR-Guided Programmable Self-Assembly of Artificial Virus-Like Nucleocapsids
Carlos Calcines-Cruz1, Ilya J Finkelstein2,3,4, Armando Hernandez-Garcia1
1Department of Chemistry of Biomacromolecules, Institute of Chemistry, National Autonomous University of Mexico, Mexico City C.P. 04510, Mexico.
Abstract:
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-like domain. Such improved self-assembly efficiency allows for discrimination between cognate versus noncognate DNA templates by the synthetic polypeptide. CRISPR-guided VLPs will help to develop programmable bioinspired nanomaterials with applications in biotechnology as well as viromimetic scaffolds to improve our understanding of viral self-assembly.
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