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| Status |
Public on Mar 21, 2024 |
| Title |
AMAR-seq: automated multimodal sequencing of DNA methylation, chromatin accessibility, and RNA expression with single-cell resolution |
| Organisms |
Homo sapiens; Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
Methylation profiling by high throughput sequencing
Other
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| Summary |
Parallel single-cell multimodal sequencing is the most intuitive and precise tool for cellular status research. In this study, we propose AMAR-seq to automate methylation, chromatin accessibility and RNA sequencing on a chip with single-cell precision. We validated the accuracy and robustness of AMAR-seq in comparison with standard single-omics methods. The high gene detection rate and genome coverage of AMAR-seq enabled us to establish a genome-wide gene expression regulatory atlas, implement single-cell copy number variation analysis, and construct a single-gene, single-base resolution gene expression, DNA methylation, and chromatin accessibility landscape. Applying AMAR-seq to investigate the process of mouse embryonic stem cell differentiation, we revealed the dynamic coupling of the epigenome and transcriptome which may contribute to the unravelling of the molecular mechanisms of early embryonic development. Collectively, these results illustrate that the employment of AMAR-seq can deeply and accurately establish single-cell multi-omics regulatory networks in a single-cell context in a cost-efficient and automated manner, which paves the way for incisive dissection of complex life procedures.
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| Overall design |
In total, 19 K562 cell line samples and 24 mESC samples were analyzed. We examined genome-wide DNA methylation, gene expression and chromatin accessibility profiles of 12 K562 single cells, two bulk K562 samples and 24 mESC single cells. We analysed DNA methylome of 3 K562 single cells by scBS-seq and transcriptome of 4 K562 single cells with smart-seq2 workflow, respectively. Mouse embryonic stem cells were grown on 6-cm culture dishes with MEF feeder cells and cultured in a complete medium supplemented with leukemia inhibitory factor (LIF) to prevent differentiation. To induce undirectional differentiation, mESCs were dissociated and replated onto 0.1% gelatin-coated dishes, and cultured without LIF in a feeder-free manner. We collected mESCs at day 0 and differentiated cells at day 3, day 6 day 9.
For library construction methods:
NOMe-seq (http://www.genome.org/cgi/doi/10.1101/gr.143008.112)
scNOMe-seq (https://elifesciences.org/articles/23203)
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| Contributor(s) |
Zeng X, Yang X, Zhong Z, Zhu Z, Li J, Song J, Yang C |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
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| Submission date |
Mar 18, 2024 |
| Last update date |
Mar 21, 2024 |
| Contact name |
Xi Zeng |
| E-mail(s) |
ivociel@outlook.com
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| Organization name |
Xiamen University
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| Street address |
No. 422, Siming South Road
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| City |
Xiamen |
| ZIP/Postal code |
361005 |
| Country |
China |
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| Platforms (2) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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| Samples (83)
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| Relations |
| BioProject |
PRJNA1089071 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE261788_RAW.tar |
6.3 Gb |
(http)(custom) |
TAR (of COV, TXT) |
SRA Run Selector |
| Raw data are available in SRA |
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