Enzymatic Ligation of Large Biomolecules to DNA

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Enzymatic Ligation of Large Biomolecules to DNA. / Sørensen, Rasmus Schøler; Okholm, Anders Hauge; Schaffert, David Henning; Kodal, Anne Louise Bank; Gothelf, Kurt Vesterager; Kjems, Jørgen.

I: ACS Nano, Bind 7, Nr. 9, 2013, s. 8098-8104.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Harvard

Sørensen, RS, Okholm, AH, Schaffert, DH, Kodal, ALB, Gothelf, KV & Kjems, J 2013, 'Enzymatic Ligation of Large Biomolecules to DNA', ACS Nano, bind 7, nr. 9, s. 8098-8104. https://doi.org/10.1021/nn403386f

APA

Sørensen, R. S., Okholm, A. H., Schaffert, D. H., Kodal, A. L. B., Gothelf, K. V., & Kjems, J. (2013). Enzymatic Ligation of Large Biomolecules to DNA. ACS Nano, 7(9), 8098-8104. https://doi.org/10.1021/nn403386f

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MLA

Vancouver

Author

Sørensen, Rasmus Schøler ; Okholm, Anders Hauge ; Schaffert, David Henning ; Kodal, Anne Louise Bank ; Gothelf, Kurt Vesterager ; Kjems, Jørgen. / Enzymatic Ligation of Large Biomolecules to DNA. I: ACS Nano. 2013 ; Bind 7, Nr. 9. s. 8098-8104.

Bibtex

@article{cee1bb6d384341a1ba71d376f42a4a16,
title = "Enzymatic Ligation of Large Biomolecules to DNA",
abstract = "The ability to synthesize, characterize, and manipulate DNA forms the foundation of a range of advanced disciplines including genomics, molecular biology, and biomolecular engineering. In particular for the latter field, DNA has proven useful as a structural or functional component in nanoscale self-assembled structures, antisense therapeutics, microarray diagnostics, and biosensors. Such applications frequently require DNA to be modified and conjugated to other macromolecules, including proteins, polymers, or fatty acids, in order to equip the system with properties required for a particular application. However, conjugation of DNA to large molecular components using classical chemistries often suffers from suboptimal yields. Here, we report the use of terminal deoxynucleotidyl transferase (TdT) for direct enzymatic ligation of native DNA to nucleotide triphosphates coupled to proteins and other large macromolecules. We demonstrate facile synthesis routes for a range of NTP-activated macromolecules and subsequent ligation to the 3′ hydroxyl group of oligodeoxynucleotides using TdT. The reaction is highly specific and proceeds rapidly and essentially to completion at micromolar concentrations. As a proof of principle, parallelly labeled oligonucleotides were used to produce nanopatterned DNA origami structures, demonstrating rapid and versatile incorporation of non-DNA components into DNA nanoarchitectures.",
keywords = "DNA Nanoteknologi, Biokonjugering, Enzymatiske reaktioner",
author = "S{\o}rensen, {Rasmus Sch{\o}ler} and Okholm, {Anders Hauge} and Schaffert, {David Henning} and Kodal, {Anne Louise Bank} and Gothelf, {Kurt Vesterager} and J{\o}rgen Kjems",
year = "2013",
doi = "10.1021/nn403386f",
language = "English",
volume = "7",
pages = "8098--8104",
journal = "A C S Nano",
issn = "1936-0851",
publisher = "American Chemical Society",
number = "9",

}

RIS

TY - JOUR

T1 - Enzymatic Ligation of Large Biomolecules to DNA

AU - Sørensen, Rasmus Schøler

AU - Okholm, Anders Hauge

AU - Schaffert, David Henning

AU - Kodal, Anne Louise Bank

AU - Gothelf, Kurt Vesterager

AU - Kjems, Jørgen

PY - 2013

Y1 - 2013

N2 - The ability to synthesize, characterize, and manipulate DNA forms the foundation of a range of advanced disciplines including genomics, molecular biology, and biomolecular engineering. In particular for the latter field, DNA has proven useful as a structural or functional component in nanoscale self-assembled structures, antisense therapeutics, microarray diagnostics, and biosensors. Such applications frequently require DNA to be modified and conjugated to other macromolecules, including proteins, polymers, or fatty acids, in order to equip the system with properties required for a particular application. However, conjugation of DNA to large molecular components using classical chemistries often suffers from suboptimal yields. Here, we report the use of terminal deoxynucleotidyl transferase (TdT) for direct enzymatic ligation of native DNA to nucleotide triphosphates coupled to proteins and other large macromolecules. We demonstrate facile synthesis routes for a range of NTP-activated macromolecules and subsequent ligation to the 3′ hydroxyl group of oligodeoxynucleotides using TdT. The reaction is highly specific and proceeds rapidly and essentially to completion at micromolar concentrations. As a proof of principle, parallelly labeled oligonucleotides were used to produce nanopatterned DNA origami structures, demonstrating rapid and versatile incorporation of non-DNA components into DNA nanoarchitectures.

AB - The ability to synthesize, characterize, and manipulate DNA forms the foundation of a range of advanced disciplines including genomics, molecular biology, and biomolecular engineering. In particular for the latter field, DNA has proven useful as a structural or functional component in nanoscale self-assembled structures, antisense therapeutics, microarray diagnostics, and biosensors. Such applications frequently require DNA to be modified and conjugated to other macromolecules, including proteins, polymers, or fatty acids, in order to equip the system with properties required for a particular application. However, conjugation of DNA to large molecular components using classical chemistries often suffers from suboptimal yields. Here, we report the use of terminal deoxynucleotidyl transferase (TdT) for direct enzymatic ligation of native DNA to nucleotide triphosphates coupled to proteins and other large macromolecules. We demonstrate facile synthesis routes for a range of NTP-activated macromolecules and subsequent ligation to the 3′ hydroxyl group of oligodeoxynucleotides using TdT. The reaction is highly specific and proceeds rapidly and essentially to completion at micromolar concentrations. As a proof of principle, parallelly labeled oligonucleotides were used to produce nanopatterned DNA origami structures, demonstrating rapid and versatile incorporation of non-DNA components into DNA nanoarchitectures.

KW - DNA Nanoteknologi

KW - Biokonjugering

KW - Enzymatiske reaktioner

U2 - 10.1021/nn403386f

DO - 10.1021/nn403386f

M3 - Journal article

C2 - 23927463

VL - 7

SP - 8098

EP - 8104

JO - A C S Nano

JF - A C S Nano

SN - 1936-0851

IS - 9

ER -