|
| Status |
Public on Oct 13, 2020 |
| Title |
Detection of genomic G-quadruplexes in living cells using a small artificial protein |
| Organisms |
Gallus gallus; Homo sapiens; Mus musculus |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
G-quadruplex (G4) structures formed by guanine-rich nucleic acids are implicated in essential physiological and pathological processes and serve as important drug targets. The genome-wide detection of G4s in living cells is important for exploring the functional role of G4s but has not yet been achieved due to the lack of a suitable G4 probe. Here we report an artificial 6.7 kDa G4 probe (G4P) protein that binds G4s with high affinity and specificity. We used it to capture G4s in living human, mouse, and chicken cells with the ChIP-Seq technique, yielding genome-wide landscape as well as details on the positions, frequencies, and sequence identities of G4 formation in these cells. Our results indicate that transcription is accompanied by a robust formation of G4s in genes. In human cells, we detected up to >123,000 G4P peaks, of which >1/3 had a fold increase of ≥5 and were present in >60% promoters and ~70% genes. Being much smaller than a scFv antibody (27 kDa) or even a nanobody (12-15 kDa), we expect that the G4P may find diverse applications in biology, medicine, and molecular devices as a G4 affinity agent.
|
| |
|
| Overall design |
We engineered a divalent G-quadruplex (G4) probe (G4P) protein that binds to the two terminal G-quartets of a G4 with high affinity and specificity through a synergy of two independent binding activities. G4-ChIP technique was developed by expressing the G4P to capture the formation of G4s in live animal cells to reveal the locations, magnitudes, and sequence identities of G4s genome-wide.
|
| |
|
| Contributor(s) |
Zheng K, Zhang J, He Y, Gong J, Wen C, Chen J, Hao Y, Zhao Y, Tan Z |
| Citation(s) |
33045726 |
| |
| Submission date |
Jun 26, 2019 |
| Last update date |
May 02, 2023 |
| Contact name |
Zheng Tan |
| E-mail(s) |
z.tan@ioz.ac.cn
|
| Organization name |
Institute of Zoology, Chinese Academy of Sciences
|
| Street address |
1 Beichen West Road, Chaoyang District
|
| City |
Beijing |
| ZIP/Postal code |
100101 |
| Country |
China |
| |
|
| Platforms (3) |
|
| Samples (18)
|
|
| Relations |
| BioProject |
PRJNA551290 |
| SRA |
SRP212149 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE133379_293T-G4P-hg19-ratio-rep1.bigWig |
321.8 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_293T-G4P-hg19-ratio-rep2.bigWig |
291.9 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_293T-G4P-hg19-rep1.narrowPeak.gz |
993.7 Kb |
(ftp)(http) |
NARROWPEAK |
| GSE133379_293T-G4P-hg19-rep2.narrowPeak.gz |
747.1 Kb |
(ftp)(http) |
NARROWPEAK |
| GSE133379_293T-G4P-hg19-subtract-rep1.bigWig |
325.4 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_293T-G4P-hg19-subtract-rep2.bigWig |
260.6 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_3T3-G4P-mm9-ratio-rep1.bigWig |
235.5 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_3T3-G4P-mm9-rep1.narrowPeak.gz |
1.0 Mb |
(ftp)(http) |
NARROWPEAK |
| GSE133379_3T3-G4P-mm9-subtract-rep1.bigWig |
239.4 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_A549-G4P-hg19-ratio-rep1.bigWig |
265.5 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_A549-G4P-hg19-ratio-rep2.bigWig |
290.3 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_A549-G4P-hg19-rep1.narrowPeak.gz |
2.9 Mb |
(ftp)(http) |
NARROWPEAK |
| GSE133379_A549-G4P-hg19-rep2.narrowPeak.gz |
3.7 Mb |
(ftp)(http) |
NARROWPEAK |
| GSE133379_A549-G4P-hg19-subtract-rep1.bigWig |
270.1 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_A549-G4P-hg19-subtract-rep2.bigWig |
293.7 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_DF-1-G4P-gal5-ratio-rep1.bigWig |
126.2 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_DF-1-G4P-gal5-rep1.narrowPeak.gz |
824.3 Kb |
(ftp)(http) |
NARROWPEAK |
| GSE133379_DF-1-G4P-gal5-subtract-rep1.bigWig |
118.0 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_H1975-G4P-hg19-ratio-rep1.bigWig |
254.1 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_H1975-G4P-hg19-ratio-rep2.bigWig |
267.2 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_H1975-G4P-hg19-rep1.narrowPeak.gz |
3.3 Mb |
(ftp)(http) |
NARROWPEAK |
| GSE133379_H1975-G4P-hg19-rep2.narrowPeak.gz |
3.6 Mb |
(ftp)(http) |
NARROWPEAK |
| GSE133379_H1975-G4P-hg19-subtract-rep1.bigWig |
232.5 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_H1975-G4P-hg19-subtract-rep2.bigWig |
272.0 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_HeLa-S3-G4P-hg19-ratio-rep1.bigWig |
284.2 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_HeLa-S3-G4P-hg19-rep1.narrowPeak.gz |
475.8 Kb |
(ftp)(http) |
NARROWPEAK |
| GSE133379_HeLa-S3-G4P-hg19-subtract-rep1.bigWig |
289.3 Mb |
(ftp)(http) |
BIGWIG |
| GSE133379_PQS-gal5.BigBed |
17.7 Mb |
(ftp)(http) |
BIGBED |
| GSE133379_PQS-hg19.BigBed |
58.7 Mb |
(ftp)(http) |
BIGBED |
| GSE133379_PQS-mm9.BigBed |
57.8 Mb |
(ftp)(http) |
BIGBED |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
Less...
More...