Quantification of cellular uptake of DNA nanostructures by qPCR

Anders Hauge Okholm; Jesper Sejrup Nielsen; Mathias Vinther; Rasmus Schøler Sørensen; David Schaffert; Jørgen Kjems

doi:10.1016/j.ymeth.2014.01.013

Quantification of cellular uptake of DNA nanostructures by qPCR

Anders Hauge Okholm, Jesper Sejrup Nielsen, Mathias Vinther, Rasmus Schøler Sørensen, David Schaffert, Jørgen Kjems

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

56 Citations (Scopus)

Abstract

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

Original language	English
Journal	Methods
Volume	67
Issue	2
Pages (from-to)	193-197
Number of pages	5
ISSN	1046-2023
DOIs	https://doi.org/10.1016/j.ymeth.2014.01.013
Publication status	Published - 15 May 2014

Access to Document

10.1016/j.ymeth.2014.01.013

Cite this

@article{4524bdaadb94430d883b6aa0b26384e1,

title = "Quantification of cellular uptake of DNA nanostructures by qPCR",

abstract = "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",

author = "Okholm, {Anders Hauge} and Nielsen, {Jesper Sejrup} and Mathias Vinther and S{\o}rensen, {Rasmus Sch{\o}ler} and David Schaffert and J{\o}rgen Kjems",

year = "2014",

month = may,

day = "15",

doi = "10.1016/j.ymeth.2014.01.013",

language = "English",

volume = "67",

pages = "193--197",

journal = "Methods",

issn = "1046-2023",

publisher = "Academic Press",

number = "2",

}

TY - JOUR

T1 - Quantification of cellular uptake of DNA nanostructures by qPCR

AU - Okholm, Anders Hauge

AU - Nielsen, Jesper Sejrup

AU - Vinther, Mathias

AU - Sørensen, Rasmus Schøler

AU - Schaffert, David

AU - Kjems, Jørgen

PY - 2014/5/15

Y1 - 2014/5/15

N2 - 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

AB - 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

U2 - 10.1016/j.ymeth.2014.01.013

DO - 10.1016/j.ymeth.2014.01.013

M3 - Journal article

C2 - 24472874

SN - 1046-2023

VL - 67

SP - 193

EP - 197

JO - Methods

JF - Methods

IS - 2

ER -

Quantification of cellular uptake of DNA nanostructures by qPCR

Abstract

Access to Document

Fingerprint

Cite this