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Sequencing abasic sites in DNA at single-nucleotide resolution.

Accepted version
Peer-reviewed

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Type

Article

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Authors

Liu, Zheng J 
Martínez Cuesta, Sergio 
van Delft, Pieter 
Balasubramanian, Shankar  ORCID logo  https://orcid.org/0000-0002-0281-5815

Abstract

In DNA, the loss of a nucleobase by hydrolysis generates an abasic site. Formed as a result of DNA damage, as well as a key intermediate during the base excision repair pathway, abasic sites are frequent DNA lesions that can lead to mutations and strand breaks. Here we present snAP-seq, a chemical approach that selectively exploits the reactive aldehyde moiety at abasic sites to reveal their location within DNA at single-nucleotide resolution. Importantly, the approach resolves abasic sites from other aldehyde functionalities known to exist in genomic DNA. snAP-seq was validated on synthetic DNA and then applied to two separate genomes. We studied the distribution of thymine modifications in the Leishmania major genome by enzymatically converting these modifications into abasic sites followed by abasic site mapping. We also applied snAP-seq directly to HeLa DNA to provide a map of endogenous abasic sites in the human genome.

Description

Keywords

Aldehydes, Base Sequence, DNA, DNA Damage, DNA-(Apurinic or Apyrimidinic Site) Lyase, Gene Knockdown Techniques, Genome, HeLa Cells, Humans, Leishmania major, Molecular Probes, Sequence Analysis, DNA, Thymine, Uracil-DNA Glycosidase

Journal Title

Nat Chem

Conference Name

Journal ISSN

1755-4330
1755-4349

Volume Title

11

Publisher

Springer Science and Business Media LLC

Rights

All rights reserved
Sponsorship
Wellcome Trust (099232/Z/12/Z)
Wellcome Trust (209441/Z/17/Z)
Wellcome Trust (grant no. 209441/Z/17/Z) Pembroke College, Cambridge and the Herchel Smith Fund Marie Curie Fellow of the European Union (grant no. FP7-PEOPLE-2013-IEF/624885)