Quantification of cellular uptake of DNA nanostructures by qPCR.
DNA nanostructures facilitating drug delivery are likely soon to be realized. In the past few decades programmed self-assembly of DNA building blocks have successfully been employed to construct sophisticated nanoscale objects. By conjugating functionalities to DNA, other molecules such as peptides, proteins and polymers can be precisely positioned on DNA nanostructures. This exceptional ability to produce modular nanoscale devices with tunable and controlled behavior has initiated an interest in employing DNA nanostructures for drug delivery. However, to obtain this the relationship between cellular interactions and structural and functional features of the DNA delivery device must be thoroughly investigated. Here, we present a rapid and robust method for the precise quantification of the component materials of DNA origami structures capable of entering cells in vitro. The quantification is performed by quantitative polymerase chain reaction, allowing a linear dynamic range of detection of five orders of magnitude. We demonstrate the use of this method for high-throughput screening, which could prove efficient to identify key features of DNA nanostructures enabling cell penetration. The method described here is suitable for quantification of in vitro uptake studies but should easily be extended to quantify DNA nanostructures in blood or tissue samples.
Interdisciplinary Nanoscience Center, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Centre for DNA Nanotechnology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark.
Anders Hauge
AH
Okholm
Interdisciplinary Nanoscience Center, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark.
Jesper Sejrup
JS
Nielsen
Interdisciplinary Nanoscience Center, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Centre for DNA Nanotechnology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark.
Mathias
M
Vinther
Interdisciplinary Nanoscience Center, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Centre for DNA Nanotechnology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark.
Rasmus Schøler
RS
Sørensen
Interdisciplinary Nanoscience Center, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Centre for DNA Nanotechnology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark.
David
D
Schaffert
Interdisciplinary Nanoscience Center, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Centre for DNA Nanotechnology, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark. Electronic address: jk@mb.au.dk.
Jørgen
J
Kjems
D017104:Bacteriophage M13 / Q000235:genetics; D002138:Calibration; D045744:Cell Line, Tumor; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004337:Drug Carriers / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D061925:Immobilized Nucleic Acids / Q000737:chemistry / Q000648:ultrastructure; D018625:Microscopy, Atomic Force; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D009690:Nucleic Acid Conformation; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection
D004279:DNA, Viral; D004337:Drug Carriers; D061925:Immobilized Nucleic Acids
Cell penetration; DNA nanostructure; Drug delivery; High-throughput; Quantification; qPCR