NU 2021-043
INVENTORS
Chad A. Mirkin*
Chi Huang
BACKGROUND
The use of CRISPR/Cas9 systems in genome editing has been limited by the inability to efficiently deliver the key editing components to tissues and across cell membranes. Common approaches to enhance tissue targeting include utilization of plasmids, transfection agents, and cationic delivery carriers. However, these approaches often suffer from insufficient delivery, uncontrollable dosing, off-target effects, and immunogenicity. Therefore, a cell-permeable, proteolytically stable, transfection agent-free, self-contained delivery system for CRISPR is needed.
ABSTRACT
Spherical nucleic acids (SNAs) developed by the Mirkin group at Northwestern are nanostructures that provide privileged cell and tissue entry. Herein, the Mirkin group developed a new class of CRISPR-SNAs named Cas9 ProSNAs as means of facilitating gene editing. Specifically, Cas9 cores are densely modified with DNA on their exterior to enhance cellular entry and preloaded with single guide RNA for genome targeting. The Cas9 proteins are fused with a GALA peptide at N-terminus and nuclear targeting signals at C-terminus to maximize endosomal escape and nuclear localization. Cas9 ProSNAs exhibit superior resistance to protease degradation and promising biocompatibility in multiple cell lines, achieving a genome editing efficiency between 32 and 47%. These new CRISPR-SNA constructs demonstrate great potential to broaden the scope of use and impact of CRISPR-Cas9 genome editing systems.
APPLICATIONS
ADVANTAGES
PUBLICATION
Huang, C. et al. (2022) CRISPER Spherical Nucleic Acids. Journal of American Chemical Society, 144(41), 18756-18760.
IP STATUS
Nationalization in US, Europe, China, Japan, South Korea, Australia, and Canada has been filed.
CRISPR SNAs enter cells and locate to nuclei for genome editing.