Tet-mediated covalent labelling of 5-methylcytosine for its genome-wide detection and sequencing

Liang Zhang, Keith E. Szulwach, Gary C. Hon, Chun Xiao Song, Beomseok Park, Miao Yu, Xingyu Lu, Qing Dai, Xiao Wang, Craig R. Street, Huiping Tan, Jung Hyun Min, Bing Ren, Peng Jin, Chuan He

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

5-methylcytosine is an epigenetic mark that affects a broad range of biological functions in mammals. The chemically inert methyl group prevents direct labelling for subsequent affinity purification and detection. Therefore, most current approaches for the analysis of 5-methylcytosine still have limitations of being either density-biased, lacking in robustness and consistency, or incapable of analysing 5-methylcytosine specifically. Here we present an approach, TAmC-Seq, which selectively tags 5-methylcytosine with an azide functionality that can be further labelled with a biotin for affinity purification, detection and genome-wide mapping. Using this covalent labelling approach, we demonstrate high sensitivity and specificity for known methylated loci, as well as increased CpG dinucleotide coverage at lower sequencing depth as compared with antibody-based enrichment, providing an improved efficiency in the 5-methylcytosine enrichment and genome-wide profiling.

Original languageEnglish (US)
Article number1517
JournalNature Communications
Volume4
DOIs
StatePublished - Mar 11 2013

Fingerprint

5-Methylcytosine
sequencing
genome
purification
Labeling
marking
affinity
Genes
Genome
biotin
mammals
loci
antibodies
Purification
sensitivity
Mammals
Azides
Chromosome Mapping
Biotin
Epigenomics

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

Tet-mediated covalent labelling of 5-methylcytosine for its genome-wide detection and sequencing. / Zhang, Liang; Szulwach, Keith E.; Hon, Gary C.; Song, Chun Xiao; Park, Beomseok; Yu, Miao; Lu, Xingyu; Dai, Qing; Wang, Xiao; Street, Craig R.; Tan, Huiping; Min, Jung Hyun; Ren, Bing; Jin, Peng; He, Chuan.

In: Nature Communications, Vol. 4, 1517, 11.03.2013.

Research output: Contribution to journalArticle

Zhang, L, Szulwach, KE, Hon, GC, Song, CX, Park, B, Yu, M, Lu, X, Dai, Q, Wang, X, Street, CR, Tan, H, Min, JH, Ren, B, Jin, P & He, C 2013, 'Tet-mediated covalent labelling of 5-methylcytosine for its genome-wide detection and sequencing', Nature Communications, vol. 4, 1517. https://doi.org/10.1038/ncomms2527
Zhang, Liang ; Szulwach, Keith E. ; Hon, Gary C. ; Song, Chun Xiao ; Park, Beomseok ; Yu, Miao ; Lu, Xingyu ; Dai, Qing ; Wang, Xiao ; Street, Craig R. ; Tan, Huiping ; Min, Jung Hyun ; Ren, Bing ; Jin, Peng ; He, Chuan. / Tet-mediated covalent labelling of 5-methylcytosine for its genome-wide detection and sequencing. In: Nature Communications. 2013 ; Vol. 4.
@article{bc0612b0b8a54de79e495c0cfae7ae1a,
title = "Tet-mediated covalent labelling of 5-methylcytosine for its genome-wide detection and sequencing",
abstract = "5-methylcytosine is an epigenetic mark that affects a broad range of biological functions in mammals. The chemically inert methyl group prevents direct labelling for subsequent affinity purification and detection. Therefore, most current approaches for the analysis of 5-methylcytosine still have limitations of being either density-biased, lacking in robustness and consistency, or incapable of analysing 5-methylcytosine specifically. Here we present an approach, TAmC-Seq, which selectively tags 5-methylcytosine with an azide functionality that can be further labelled with a biotin for affinity purification, detection and genome-wide mapping. Using this covalent labelling approach, we demonstrate high sensitivity and specificity for known methylated loci, as well as increased CpG dinucleotide coverage at lower sequencing depth as compared with antibody-based enrichment, providing an improved efficiency in the 5-methylcytosine enrichment and genome-wide profiling.",
author = "Liang Zhang and Szulwach, {Keith E.} and Hon, {Gary C.} and Song, {Chun Xiao} and Beomseok Park and Miao Yu and Xingyu Lu and Qing Dai and Xiao Wang and Street, {Craig R.} and Huiping Tan and Min, {Jung Hyun} and Bing Ren and Peng Jin and Chuan He",
year = "2013",
month = "3",
day = "11",
doi = "10.1038/ncomms2527",
language = "English (US)",
volume = "4",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - Tet-mediated covalent labelling of 5-methylcytosine for its genome-wide detection and sequencing

AU - Zhang, Liang

AU - Szulwach, Keith E.

AU - Hon, Gary C.

AU - Song, Chun Xiao

AU - Park, Beomseok

AU - Yu, Miao

AU - Lu, Xingyu

AU - Dai, Qing

AU - Wang, Xiao

AU - Street, Craig R.

AU - Tan, Huiping

AU - Min, Jung Hyun

AU - Ren, Bing

AU - Jin, Peng

AU - He, Chuan

PY - 2013/3/11

Y1 - 2013/3/11

N2 - 5-methylcytosine is an epigenetic mark that affects a broad range of biological functions in mammals. The chemically inert methyl group prevents direct labelling for subsequent affinity purification and detection. Therefore, most current approaches for the analysis of 5-methylcytosine still have limitations of being either density-biased, lacking in robustness and consistency, or incapable of analysing 5-methylcytosine specifically. Here we present an approach, TAmC-Seq, which selectively tags 5-methylcytosine with an azide functionality that can be further labelled with a biotin for affinity purification, detection and genome-wide mapping. Using this covalent labelling approach, we demonstrate high sensitivity and specificity for known methylated loci, as well as increased CpG dinucleotide coverage at lower sequencing depth as compared with antibody-based enrichment, providing an improved efficiency in the 5-methylcytosine enrichment and genome-wide profiling.

AB - 5-methylcytosine is an epigenetic mark that affects a broad range of biological functions in mammals. The chemically inert methyl group prevents direct labelling for subsequent affinity purification and detection. Therefore, most current approaches for the analysis of 5-methylcytosine still have limitations of being either density-biased, lacking in robustness and consistency, or incapable of analysing 5-methylcytosine specifically. Here we present an approach, TAmC-Seq, which selectively tags 5-methylcytosine with an azide functionality that can be further labelled with a biotin for affinity purification, detection and genome-wide mapping. Using this covalent labelling approach, we demonstrate high sensitivity and specificity for known methylated loci, as well as increased CpG dinucleotide coverage at lower sequencing depth as compared with antibody-based enrichment, providing an improved efficiency in the 5-methylcytosine enrichment and genome-wide profiling.

UR - http://www.scopus.com/inward/record.url?scp=84874607005&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84874607005&partnerID=8YFLogxK

U2 - 10.1038/ncomms2527

DO - 10.1038/ncomms2527

M3 - Article

VL - 4

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

M1 - 1517

ER -