Gene-encoding DNA origami for mammalian cell expression

• Published in: Nature communications Vol. 14; no. 1; p. 1017
• Main Authors: Kretzmann, Jessica A., Liedl, Anna, Monferrer, Alba, Mykhailiuk, Volodymyr, Beerkens, Samuel, Dietz, Hendrik
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 23.02.2023; Nature Publishing Group; Nature Portfolio
• Subjects: 101/28; 13/106; 13/109; 13/31; 14/63; 38/35; 38/44; 631/61; 639/925/926; Deoxyribonucleic acid; DNA; DNA - chemistry; DNA - genetics; Gene expression; Gene transfer; Genes; Humanities and Social Sciences; Mammalian cells; Mammals; multidisciplinary; Nanostructures - chemistry; Nanotechnology; Nucleic Acid Conformation; Nucleotide sequence; Scaffolds; Science; Science (multidisciplinary); Tissue engineering
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-023-36601-1

DNA origami may enable more versatile gene delivery applications through its ability to create custom nanoscale objects with specific targeting, cell-invading, and intracellular effector functionalities. Toward this goal here we describe the expression of genes folded in DNA origami objects delivered to mammalian cells. Genes readily express from custom-sequence single-strand scaffolds folded within DNA origami objects, provided that the objects can denature in the cell. We demonstrate enhanced gene expression efficiency by including and tuning multiple functional sequences and structures, including virus-inspired inverted-terminal repeat-like (ITR) hairpin motifs upstream or flanking the expression cassette. We describe gene-encoding DNA origami bricks that assemble into multimeric objects to enable stoichiometrically controlled co-delivery and expression of multiple genes in the same cells. Our work provides a framework for exploiting DNA origami for gene delivery applications. DNA origami may enable more versatile gene delivery applications through its ability to create custom nanoscale objects. Here the authors show that genes folded in DNA origami with custom scaffolds express efficiently when delivered to mammalian cells and can be assembled into multimeric arrays to deliver and express defined ratios of multiple genes simultaneously.