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E-GEOD-64115 - Deterministic direct reprogramming of somatic cells to pluripotency [RRBS]

Status
Released on 13 December 2014, last updated on 2 January 2015
Organism
Mus musculus
Samples (8)
Protocols (4)
Description
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. 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. 
Experiment type
methylation profiling by high throughput sequencing 
Contacts
Asaf Zviran <azviran@gmail.com>, E Chomsky, J H Hanna, S Mukamel, S Shipony, Y Rais
MINSEQE
Exp. designProtocolsVariablesProcessedSeq. reads
Files
Investigation descriptionE-GEOD-64115.idf.txt
Sample and data relationshipE-GEOD-64115.sdrf.txt
Processed data (2)E-GEOD-64115.processed.1.zip, E-GEOD-64115.processed.2.zip
Links