DNA Cage Delivery to Mammalian Cells

Abstract

DNA cages are nanometer-scale polyhedral structures formed by self-assembly from synthetic DNA oligonucleotides. Potential applications include in vivo imaging and the targeted delivery of macromolecules into living cells. We report an investigation of the ability of a model cage, a DNA tetrahedron, to enter live cultured mammalian cells. Cultured human embryonic kidney cells were treated with a range of fluorescently labeled DNA tetrahedra and subsequently examined using confocal microscopy and flow cytometry. Substantial uptake of tetrahedra into cells was observed both when the cells were treated with tetrahedra alone and when the cells were treated with a mixture of tetrahedra and a transfection reagent. Analysis of the subcellular localization of transfected tetrahedra using confocal microscopy and organelle staining indicates that the cages are located in the cytoplasm. FRET experiments indicate that the DNA cages remain substantially intact within the cells for at least 48 h after transfection. This is a first step toward the use of engineered DNA nanostructures to deliver and control the activity of cargoes within cells.

Structural insights on biologically relevant cationic membranes by ESR spectroscopy

“…DNA has been extensively employed as a biotechnological tool, with applications in different forms of genetic modification, such as gene therapy and silencing (Fichou and Gersbach 2016), in vaccine development (Aps et al 2016) and as vaccine adjuvants (Rozenfeld et al 2012), and even in the delivery of macromolecules (as hydrogels and DNA cages) (Campolongo et al 2010;Walsh et al 2011). In many of these applications, the DNA molecules used are either in the form of long and circular plasmids (see, for example, Aps et al 2016;Robinson-Hamm and Gersbach 2016) or as small linear oligonucleotides (Fichou and Férec 2006;Rozenfeld et al 2012).…”

Temperature-Controlled Encapsulation and Release of an Active Enzyme in the Cavity of a Self-Assembled DNA Nanocage

“…34 More recently DNA origami nanoarrays were demonstrated stable in cell lysates. 35 DNA cage structures have also been demonstrated capable of entering live mammalian cells, 36 while DNA tubes or barrels have been demonstrated directable to specific cell types to which they can either deliver 37 or expose 31 their specific cargo.…”

Supramolecular Wireframe DNA Polyhedra: Assembly and Applications

“…[72][73][74] Turberfield et al found that DNA tetrahedra could enter a mammalian cell with or without the need of a transfection reagent. The tetrahedral DNA cage remained its structural intactness in the cellular environment for at least 48 h. [75] Fan et al investigated the endocytotic internalization of a DNA tetrahedron and its subsequent intracellular transport. [76] The DNA nanocages mainly resided in the lysosomes after a rapid internalization, which would affect their functions as a delivery agent.…”