Transcription of click-linked DNA in human cells
Transcription of click-linked DNA in human cells
Click DNA ligation promises an alternative to the current enzymatic approaches for DNA assembly, with the ultimate goal of using efficient chemical reactions for the total chemical synthesis and assembly of genes and genomes. Such an approach would enable the incorporation of various chemically modified bases throughout long stretches of DNA, a feat not possible with current polymerase-based methods. An unequivocal requirement for this approach is the biocompatibility of the resulting triazole-linked DNA. The correct function of this unnatural DNA linker in human cells is demonstrated here by using a click-linked gene encoding the fluorescent protein mCherry. Reverse transcription of mRNA isolated from these cells and subsequent sequencing of the mCherry cDNA shows error-free transcription. Nucleotide excision repair (NER) is shown to not play a role in the observed biocompatibility by using a NER-deficient human cell line. This is the first example of a non-natural DNA linker being functional in a eukaryotic cell
click chemistry, dna ligation, gene technology, nucleic acids, synthetic biology
2362-2365
Birts, Charles N.
8689ddad-ba47-4ca6-82c5-001315dbd250
Sanzone, A. Pia
e57b6591-2c10-4abc-b398-f32ee0e4e096
El-Sagheer, Afaf H.
05b8295a-64ad-4fdf-ad57-c34934a46c04
Blaydes, Jeremy P.
e957f999-fd91-4f77-ad62-5b4ef069b15b
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Tavassoli, Ali
d561cf8f-2669-46b5-b6e1-2016c85d63b2
24 February 2014
Birts, Charles N.
8689ddad-ba47-4ca6-82c5-001315dbd250
Sanzone, A. Pia
e57b6591-2c10-4abc-b398-f32ee0e4e096
El-Sagheer, Afaf H.
05b8295a-64ad-4fdf-ad57-c34934a46c04
Blaydes, Jeremy P.
e957f999-fd91-4f77-ad62-5b4ef069b15b
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Tavassoli, Ali
d561cf8f-2669-46b5-b6e1-2016c85d63b2
Birts, Charles N., Sanzone, A. Pia, El-Sagheer, Afaf H., Blaydes, Jeremy P., Brown, Tom and Tavassoli, Ali
(2014)
Transcription of click-linked DNA in human cells.
Angewandte Chemie International Edition, 53 (9), .
(doi:10.1002/anie.201308691).
(PMID:24452865)
Abstract
Click DNA ligation promises an alternative to the current enzymatic approaches for DNA assembly, with the ultimate goal of using efficient chemical reactions for the total chemical synthesis and assembly of genes and genomes. Such an approach would enable the incorporation of various chemically modified bases throughout long stretches of DNA, a feat not possible with current polymerase-based methods. An unequivocal requirement for this approach is the biocompatibility of the resulting triazole-linked DNA. The correct function of this unnatural DNA linker in human cells is demonstrated here by using a click-linked gene encoding the fluorescent protein mCherry. Reverse transcription of mRNA isolated from these cells and subsequent sequencing of the mCherry cDNA shows error-free transcription. Nucleotide excision repair (NER) is shown to not play a role in the observed biocompatibility by using a NER-deficient human cell line. This is the first example of a non-natural DNA linker being functional in a eukaryotic cell
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e-pub ahead of print date: 22 January 2014
Published date: 24 February 2014
Keywords:
click chemistry, dna ligation, gene technology, nucleic acids, synthetic biology
Organisations:
Cancer Sciences
Identifiers
Local EPrints ID: 362373
URI: http://eprints.soton.ac.uk/id/eprint/362373
ISSN: 1433-7851
PURE UUID: 98d73c33-c0cd-48a9-b818-31e5caf3816e
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Date deposited: 24 Feb 2014 12:30
Last modified: 15 Mar 2024 03:26
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Author:
A. Pia Sanzone
Author:
Afaf H. El-Sagheer
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