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| Status |
Public on Jun 12, 2017 |
| Title |
Human genome wide repair map of DNA damage caused by the cigarette smoke carcinogen benzo[a]pyrene |
| Organism |
Homo sapiens |
| Experiment type |
Other
|
| Summary |
Benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon, is the major cause of lung cancer. It forms covalent DNA adducts after metabolic activation and induces mutations. We have developed a method capturing oligonucleotides carrying bulky base adducts, including UV-induced cyclobutane pyrimidine dimers (CPDs) and BaP diol epoxide-deoxyguanosine (BPDE-dG), which are removed from the genome by nucleotide excision repair. The isolated oligonucleotides are ligated to adaptors and after damage-specific immunoprecipitation the adaptor-ligated oligonucleotides are converted to dsDNA with an appropriate translesion DNA synthesis (TLS) polymerase followed by PCR amplification and next-generation sequencing (NGS) to generate genome-wide repair maps. We have named this method as translesion eXcision Repair-sequencing (tXR-seq). In contrast to our previously described XR-seq method, the tXR-seq does not depend on repair/removal of the damage in the excised oligonucleotides and hence it is applicable to essentially all DNA damages processed by nucleotide excision repair. Here, we present the excision repair maps for CPDs and BPDE-dG adducts generated by tXR-Seq for the human genome. Additionally, we observe novel sequence specificity of BPDE-dG excision repair by using tXR-seq.
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| Overall design |
We performed tXR-seq for UV-induced CPD and BPDE-dG repair in GM12878 cell line. We also compared our tXR-seq data with our published dataset from XR-seq for CPD and (6-4)PP (GEO accession: GSE67941).
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| Contributor(s) |
Li W, Hu J, Adebal O, Adar S, Yang Y, Chiou Y, Sancar A |
| Citation(s) |
28607059, 39933696 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 ES027255 |
Single Nucleotide Resolution Map of Formation and Repair of Bulky Adducts in the Human Genome |
UNIVERSITY OF NORTH CAROLINA AT CHAPEL HILL |
AZIZ SANCAR |
| R35 GM118102 |
Molecular Mechanism of Mammalian DNA Excision Repair, DNA Damage Checkpoints and the Circadian Clock |
UNIVERSITY OF NORTH CAROLINA AT CHAPEL HILL |
AZIZ SANCAR |
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| |
| Submission date |
Apr 12, 2017 |
| Last update date |
Feb 25, 2025 |
| Contact name |
Wentao Li |
| Organization name |
University of Georgia
|
| Department |
Environmental Health Science
|
| Lab |
Li, Wentao
|
| Street address |
500 D. W Brooks Drive
|
| City |
Athens |
| State/province |
GA |
| ZIP/Postal code |
30602 |
| Country |
USA |
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| Platforms (2) |
| GPL11154 |
Illumina HiSeq 2000 (Homo sapiens) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA382636 |
| SRA |
SRP103822 |
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