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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.
Nano Letters
|March 17, 2021
Summary
Researchers used CRISPR-Cas12a to program synthetic polypeptides for self-assembly into virus-like particles (VLPs) on specific DNA. This breakthrough enables precise control over gene delivery systems and biomaterials.
Area of Science:
- Biotechnology
- Molecular Biology
- Materials Science
Background:
- Designer proteins inspire advanced gene-delivery systems and models for viral assembly.
- Self-assembly of engineered proteins on nucleic acid templates, a key viral trait, remains challenging.
Purpose of the Study:
- To develop a programmable method for self-assembling synthetic polypeptides into virus-like particles (VLPs) on specific DNA templates.
- To leverage CRISPR-Cas12a programmability to mimic viral packaging mechanisms.
Main Methods:
- Utilized nuclease-dead Cas12a (dCas12a) proteins positioned on DNA templates to direct polypeptide self-assembly.
- Engineered dCas12a for enhanced particle growth by dimerization with a silk-like domain.
- Investigated self-assembly efficiency and DNA encapsidation.
Main Results:
- Successfully directed the nucleation and growth of synthetic polypeptides into VLPs on specific DNA molecules using CRISPR-guided dCas12a.
- Achieved full DNA encapsidation on a 48.5 kbp template at limiting polypeptide concentrations.
- Demonstrated improved self-assembly efficiency and template discrimination with engineered dCas12a.
Conclusions:
- CRISPR-guided VLP assembly offers a programmable approach for creating bioinspired nanomaterials.
- This method enhances understanding of viral self-assembly and opens avenues for advanced biotechnology applications.
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