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| Status |
Public on Dec 12, 2010 |
| Title |
Selective chemical labeling reveals intragenic enrichment of 5-hydroxymethylcytosine in mouse cerebellum |
| Organism |
Mus musculus |
| Experiment type |
Methylation profiling by high throughput sequencing
|
| Summary |
5-Methylcytosine (5-mC) is an important DNA modification found in eukaryotes that impacts gene regulation and disease pathogenesis. Recently, 5-hydroxymethylcytosine (5-hmC), another form of DNA modification, has been identified in substantial amounts in certain mammalian cell types; however, its roles as well as its distribution in mammalian genomes are unknown. Here we present a selective chemical labeling method for 5-hmC by utilizing T4 bacteriophage BGT-glucosyltransferase to transfer an engineered glucose moiety containing an azide group onto the hydroxyl group of 5-hmC, which in turn can chemically incorporate a biotin group for detection, affinity enrichment, and sequencing of 5-hmC in mammalian genomes. Using this highly effective method, we demonstrate that 5-hmC is present in human cell lines beyond those previously recognized. We also find a gene expression level-dependent enrichment of intragenic 5-hmC in mouse cerebellum and an age-dependent acquisition of this modification in specific gene bodies linked to neurodegenerative disorders
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| Overall design |
Identification of 5hmC enriched genmoic regions in mouse cerebellum
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| |
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| Contributor(s) |
Peng Jin CH, Szulwach KE |
| Citation(s) |
21151123 |
| Submission date |
Nov 15, 2010 |
| Last update date |
May 15, 2019 |
| Contact name |
Keith E Szulwach |
| E-mail(s) |
kszulwa@emory.edu
|
| Phone |
404-712-0796
|
| Organization name |
Emory University
|
| Department |
Human Genetics
|
| Lab |
Jin Lab
|
| Street address |
615 Michael Street, Whitehead Suite 301
|
| City |
Atlanta |
| State/province |
GA |
| ZIP/Postal code |
30322 |
| Country |
USA |
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| Platforms (1) |
| GPL9250 |
Illumina Genome Analyzer II (Mus musculus) |
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| Samples (13)
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| Relations |
| SRA |
SRP004445 |
| BioProject |
PRJNA134133 |
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