Gene-encoding DNA origami for mammalian cell expression

Area of Science:

• Nanotechnology
• Molecular Biology
• Gene Therapy

Background:

• DNA origami allows for the creation of custom nanoscale objects.
• Gene delivery is crucial for various therapeutic and research applications.
• Current gene delivery methods face limitations in efficiency and control.

Purpose of the Study:

• To demonstrate the expression of genes from DNA origami objects in mammalian cells.
• To enhance gene expression efficiency using optimized DNA origami designs.
• To enable controlled co-delivery and expression of multiple genes using DNA origami.

Main Methods:

• Designing and folding single-strand DNA scaffolds into custom origami objects encoding genes.
• Delivering DNA origami objects into mammalian cells.
• Optimizing expression cassettes with functional sequences, including inverted terminal repeat (ITR) motifs.
• Assembling gene-encoding DNA origami bricks into multimeric objects for co-delivery.

Main Results:

• Genes successfully expressed from DNA origami objects delivered to mammalian cells.
• Gene expression efficiency was significantly enhanced by incorporating and tuning functional sequences and structures.
• Virus-inspired ITR hairpin motifs improved gene expression.
• Multimeric DNA origami objects enabled stoichiometrically controlled co-delivery and expression of multiple genes.

Conclusions:

• DNA origami is a versatile platform for gene delivery applications.
• Optimized DNA origami designs can achieve efficient gene expression in mammalian cells.
• This framework supports the development of advanced gene delivery systems using DNA nanotechnology.