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| Status |
Public on Dec 13, 2014 |
| Title |
Deterministic direct reprogramming of somatic cells to pluripotency [RRBS] |
| Organism |
Mus musculus |
| Experiment type |
Methylation profiling by high throughput sequencing
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| Summary |
Somatic cells can be directly reprogrammed to pluripotency by exogenous expression of transcription factors, classically Oct4, Sox2, Klf4 and c-Myc. While distinct types of somatic cells can be reprogramed with varying efficiencies and by different modified reprogramming protocols, induced pluripotent stem cell (iPSC) induction remains inefficient and stochastic where a fraction of the cells converts into iPSCs. The nature of rate limiting barrier(s) preventing majority of cells to convert into iPSCs remains elusive. Here we show that neutralizing Mbd3, a core member of the Mbd3/NURD co-repressor and chromatin-remodeling complex, results in deterministic and synchronized reprogramming of multiple differentiated cell types to pluripotency. 100% of Mbd3 depleted mouse and human somatic cells convert into iPSCs after seven days of reprogramming factor induction. Our findings delineate a critical pathway blocking the reestablishment of pluripotency, and offer a novel platform for future dissection of epigenetic dynamics leading to iPSC formation at high resolution.
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| Overall design |
Reduced representation bisulfite sequencing (RRBS) was applied to mouse iPS cells and mouse embryonic fibroblast (MEF) before and after DOX induction (initiating reprogramming by OSKM factors) from randomly selected Mbd3+/+ and Mbd3flox/- clonal cell line series. Polyclonal donor cell cultures were harvested at days 0,4 and 8 after DOX reprogramming without selection or sorting for any marker or passaging, and mapped for similarity to subcloned iPSC lines.
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| Contributor(s) |
Rais Y, Chomsky E, Shipony S, Mukamel S, Hanna JH |
| Citation(s) |
25830885 |
| Submission date |
Dec 12, 2014 |
| Last update date |
May 15, 2019 |
| Contact name |
Asaf Zviran |
| E-mail(s) |
azviran@gmail.com
|
| Organization name |
Weizmann Institute of Science
|
| Department |
Molecular Genetics
|
| Lab |
Hanna's Laboratory for Pluripotent Cell Studies
|
| Street address |
234 Herzl Street
|
| City |
Rehovot |
| ZIP/Postal code |
76100 |
| Country |
Israel |
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| Platforms (1) |
| GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
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| Samples (8)
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| GSM1564675 |
RRBS of wild-type MEF cells before DOX induction |
| GSM1564676 |
RRBS of wild-type MEF cells at day 4 after DOX induction |
| GSM1564677 |
RRBS of wild-type MEF cells at day 8 after DOX induction |
| GSM1564678 |
RRBS of wild-type iPSCs |
| GSM1564679 |
RRBS of Mbd3f/- MEF cells before DOX induction |
| GSM1564680 |
RRBS of Mbd3f/- MEF cells at day 4 after DOX induction |
| GSM1564681 |
RRBS of Mbd3f/- MEF cells at day 8 after DOX induction |
| GSM1564682 |
RRBS of Mbd3f/- iPSCs |
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| This SubSeries is part of SuperSeries: |
| GSE49767 |
Deterministic direct reprogramming of somatic cells to pluripotency |
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| Relations |
| BioProject |
PRJNA270182 |
| SRA |
SRP051103 |
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