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GEO help: Mouse over screen elements for information. |
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| Status |
Public on May 19, 2022 |
| Title |
Cell cycle states of pluripotent cells dictate lineage decisions |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
Cell cycle and differentiation decisions are tightly linked; however, the underlying principles that drive these decisions are not fully understood. Here, we combined cell-cycle reporter system and single-cell RNA-seq profiling, to study the transcriptomes of mouse embryonic stem cells (ESCs) in the context of cell cycle states and differentiation. By applying a retinoic acid-based differentiation protocol we show that only cells in the G2/M phase are capable of differentiating into extraembryonic endoderm cells (XENs), whereas cells in the G1 phase predominantly produce epiblast stem cells (EpiSCs). Mechanistically, we show that Esrrb is a potent XEN state inducer as it is predominantly upregulated during G2/M phase, and cells engineered to overexpress Esrrb during G1 phase forced the cells to become XENs. Interestingly, this phenomenon is unique to the pluripotency ground state as sorting cells after 48 hours of differentiation resulted in a cell cycle-independent differentiation decisions. Cells in both G1 and G2/M phases contributed equally to EpiSC and XEN cellular lineages. Taken together, this study reveals an important functional link between cell-cycle states of pluripotent cells and lineage decisions, emphasizing the regulatory role of cell cycle during exit from pluripotency and can be further expand our understanding of early differentiation events.
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| Overall design |
We used R1 mESC line, each experiment includes technical duplicats.we established R1 clones with Fucci, sorted according to cell cycle stage at 0, 2 , 4 days of RA differentiation - then bulk or single cell analysis. We also used R1-fucci-Over expression ESRRB cells and zhbtc4 esrrb KO cells and preformes single cell analysis on them.
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| Contributor(s) |
Herchcovici Levy S, Arnon L, Feldman S, Alajem A, Awawdy M, Lahav S, Bavli D, Sun X, Buganim Y, Ram O |
| Citation(s) |
35594859 |
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| Submission date |
Jun 17, 2021 |
| Last update date |
Jun 30, 2022 |
| Contact name |
Muhammad Awawdy |
| E-mail(s) |
muhammad.awawdy@mail.huji.ac.il
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| Phone |
+972528482707
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| Organization name |
Hebrew university of jerusalem
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| Lab |
Muhammad Awawdy
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| Street address |
The Hebrew University, The Edmond J. Safra Campus - Givat Ram
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| City |
jerusalem |
| ZIP/Postal code |
9190401 |
| Country |
Israel |
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| Platforms (2) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
| GPL21626 |
NextSeq 550 (Mus musculus) |
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| Samples (65)
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| Relations |
| BioProject |
PRJNA738859 |
| SRA |
SRP324494 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE178390_RAW.tar |
130.0 Mb |
(http)(custom) |
TAR (of BED, RESULTS) |
| GSE178390_chipK4me1_KOES_Promoters.bed.gz |
1.6 Mb |
(ftp)(http) |
BED |
| GSE178390_chipK4me1_WTES_Promoters.bed.gz |
1.5 Mb |
(ftp)(http) |
BED |
| GSE178390_chipK4me2_KOES_Enhancers.bed.gz |
808.6 Kb |
(ftp)(http) |
BED |
| GSE178390_chipK4me2_KOES_Promoters.bed.gz |
1.8 Mb |
(ftp)(http) |
BED |
| GSE178390_chipK4me2_WTES_Enhancers.bed.gz |
808.0 Kb |
(ftp)(http) |
BED |
| GSE178390_chipK4me2_WTES_Promoters.bed.gz |
1.9 Mb |
(ftp)(http) |
BED |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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