Intracellular Delivery of a Planar DNA Origami Structure by the Transferrin‐Receptor Internalization Pathway

Abstract

DNA origami provides rapid access to easily functionalized, nanometer-sized structures making it an intriguing platform for the development of defined drug delivery and sensor systems. Low cellular uptake of DNA nanostructures is a major obstacle in the development of DNA-based delivery platforms. Herein, significant strong increase in cellular uptake in an established cancer cell line by modifying a planar DNA origami structure with the iron transport protein transferrin (Tf) is demonstrated. A variable number of Tf molecules are coupled to the origami structure using a DNA-directed, site-selective labeling technique to retain ligand functionality. A combination of confocal fluorescence microscopy and quantitative (qPCR) techniques shows up to 22-fold increased cytoplasmic uptake compared to unmodified structures and with an efficiency that correlates to the number of transferrin molecules on the origami surface.

Allosteric DNA nanoswitches for controlled release of a molecular cargo triggered by biological inputs

“…Finally, because several DNA-based vehicles and nanostructures have been associated with innovative approaches to enhance their transfection properties and overcome some of the current barriers in drug delivery, 35 our strategy might allow extension of the functionality of these devices so that they can be regulated by different biological inputs (such as antibodies and proteins) and it could thus open the door for the next generation of DNA-based drug delivery systems.…”

Multifunctional DNA Origami Nanoplatforms for Drug Delivery

“…Moreover, Tf has been successfully employed to facilitate the delivery of macromolecular payloads and various DNA constructs in target cells. Kjems and co‐workers first equipped a planar DNA origami with Tf ligand to increase cellular uptake in a cancer cell line (Figure a) . The Tf‐modified DNA origami nanostructure demonstrated efficient cellular uptake through receptor‐mediated endocytosis.…”

“…Reproduced from Ref. with permission from Wiley‐VCH, Copyright 2016. b) DNA nanorobot for antibody fragment delivery. Reproduced from Ref.…”

Advances in DNA Origami–Cell Interfaces

“…Whereas the approaches described above involve adding modifications to improve passive uptake through the cell membrane, another possible approach is to exploit pre‐existing endocytotic mechanisms to get nanostructures into the cell. Schaffert and co‐workers used transferrin‐mediated endocytosis, which is one of the most common endocytotic pathways in mammalian cells, to shuttle DNA nanostructures into the intracellular space . The authors attached the transport protein transferrin onto the edges of a planar DNA origami structure (Figure C) and observed up to a 22‐fold increase in cytoplasmic uptake compared with that of naked nanostructures.…”