PMID- 30746092
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20231005
IS  - 2041-6520 (Print)
IS  - 2041-6539 (Electronic)
IS  - 2041-6520 (Linking)
VI  - 10
IP  - 2
DP  - 2019 Jan 14
TI  - Bisulfite-free, single base-resolution analysis of 5-hydroxymethylcytosine in 
      genomic DNA by chemical-mediated mismatch.
PG  - 447-452
LID - 10.1039/c8sc04272a [doi]
AB  - 5-Hydroxymethylcytosine (5hmC) is known as one of the vital players in nuclear 
      reprogramming and the process of active DNA demethylation. Although the 
      development of whole-genome sequencing methods for modified cytosine bases has 
      burgeoned, the easily operated gene-specific loci detection of 5hmC has rarely 
      been reported. Herein, we present a single-base resolution approach, i.e., 
      chemical-assisted mismatch sequencing (CAM-Seq), which, when combined with 
      traditional oxidation and chemical labeling mediation, can be used for mapping 
      5hmC at base resolution. We employ chemical oxidation to transform 5hmC to 
      5-formylcytosine (5fC), followed by chemical labeling to induce C-to-T base 
      changes owing to the fact that the loss of the exocyclic 4-amino group of labeled 
      5fC leads to C to T conversion and subsequent pairing with adenosine (A) in PCR. 
      The feasibility of CAM-Seq is demonstrated in different synthetic oligonucleotide 
      models as well as in part of the genome of 5hmC-rich mouse embryonic stem cells 
      (mESCs). Moreover, the gene fragment containing 5hmC can be easily biotinylated 
      after oxidation, showing high enrichment efficiency. Our method has the potential 
      capability to map 5hmC in genomic DNA and thus will contribute to promoting the 
      understanding of the epigenetic modification of 5hmC.
FAU - Wang, Yafen
AU  - Wang Y
AD  - College of Chemistry and Molecular Sciences , Key Laboratory of Biomedical 
      Polymers of Ministry of Education , The Institute for Advanced Studies , Hubei 
      Province Key Laboratory of Allergy and Immunology , Wuhan University , Wuhan , 
      Hubei 430072 , P. R. China . Email: xzhou@whu.edu.cn ; ; Tel: +86-27-68756663.
FAU - Zhang, Xiong
AU  - Zhang X
AD  - College of Chemistry and Molecular Sciences , Key Laboratory of Biomedical 
      Polymers of Ministry of Education , The Institute for Advanced Studies , Hubei 
      Province Key Laboratory of Allergy and Immunology , Wuhan University , Wuhan , 
      Hubei 430072 , P. R. China . Email: xzhou@whu.edu.cn ; ; Tel: +86-27-68756663.
FAU - Wu, Fan
AU  - Wu F
AD  - College of Chemistry and Molecular Sciences , Key Laboratory of Biomedical 
      Polymers of Ministry of Education , The Institute for Advanced Studies , Hubei 
      Province Key Laboratory of Allergy and Immunology , Wuhan University , Wuhan , 
      Hubei 430072 , P. R. China . Email: xzhou@whu.edu.cn ; ; Tel: +86-27-68756663.
FAU - Chen, Zonggui
AU  - Chen Z
AD  - College of Life Science , Wuhan University , Wuhan , Hubei 430072 , P. R. China.
FAU - Zhou, Xiang
AU  - Zhou X
AUID- ORCID: 0000-0002-1829-9368
AD  - College of Chemistry and Molecular Sciences , Key Laboratory of Biomedical 
      Polymers of Ministry of Education , The Institute for Advanced Studies , Hubei 
      Province Key Laboratory of Allergy and Immunology , Wuhan University , Wuhan , 
      Hubei 430072 , P. R. China . Email: xzhou@whu.edu.cn ; ; Tel: +86-27-68756663.
LA  - eng
PT  - Journal Article
DEP - 20181011
PL  - England
TA  - Chem Sci
JT  - Chemical science
JID - 101545951
PMC - PMC6335847
EDAT- 2019/02/13 06:00
MHDA- 2019/02/13 06:01
PMCR- 2018/10/11
CRDT- 2019/02/13 06:00
PHST- 2018/09/26 00:00 [received]
PHST- 2018/10/11 00:00 [accepted]
PHST- 2019/02/13 06:00 [entrez]
PHST- 2019/02/13 06:00 [pubmed]
PHST- 2019/02/13 06:01 [medline]
PHST- 2018/10/11 00:00 [pmc-release]
AID - c8sc04272a [pii]
AID - 10.1039/c8sc04272a [doi]
PST - epublish
SO  - Chem Sci. 2018 Oct 11;10(2):447-452. doi: 10.1039/c8sc04272a. eCollection 2019 
      Jan 14.