A Versatile Nonviral Delivery System for Multiplex Gene-Editing in the Liver

Area of Science:

• Biotechnology
• Nanomedicine
• Genetic Engineering

Background:

• CRISPR technology offers potential for genetic-level therapeutic strategies.
• Efficient delivery of CRISPR components is crucial for in vivo applications.
• Targeting lipid metabolism genes could improve cardiovascular health.

Purpose of the Study:

• To develop and evaluate a liposome-coated mesoporous silica nanoparticle (lipoMSN) as a CRISPR delivery system.
• To investigate the efficacy of multiplex gene editing in the liver using lipoMSN.
• To assess the therapeutic potential of combinatorial gene editing for lipid metabolism disorders.

Main Methods:

• lipoMSN formulation for efficient loading of Cas9 plasmid or ribonucleoprotein (RNP) complexes.
• In vitro and in vivo delivery of RNPs targeting PCSK9, APOC3, and ANGPTL3 genes.
• Assessment of gene-editing efficiency and lipid metabolism changes.

Main Results:

• lipoMSN demonstrated superior gene-editing efficiency compared to Lipofectamine CRISPRMax for single-gene targets.
• Multiplex gene editing was achieved with maintained efficiency despite reduced RNP dosage.
• Combinatorial RNP delivery via lipoMSN resulted in synergistic improvements in lipid metabolism, including a 50% decrease in serum cholesterol.

Conclusions:

• lipoMSN is an effective platform for CRISPR delivery, enabling efficient multiplex gene editing in the liver.
• Combinatorial gene editing using lipoMSN offers enhanced therapeutic effects on lipid metabolism.
• This versatile platform holds significant potential for developing novel gene-editing therapeutics.