Reprogramming fibroblasts to pluripotency
Abstract/Contents
- Abstract
- Reprogramming of somatic nuclei to a pluripotent state can be achieved by two approaches: somatic cell nuclear transfer (cloning) and transcription factor transduction to generate induced pluripotent stem cells (iPS). Early molecular events in reprogramming are not well understood, in large part due to the inefficiency of nuclear transfer and iPS generation. This inefficiency has led to the hypothesis that early reprogramming is a stochastic rather than a deterministic process. We hypothesized that iPS generation could be improved by the identification of early molecular events and pathways that may enhance reprogramming. In order to model reprogramming of fibroblasts towards pluripotency, we generated mouse embryonic stem cell X human fibroblast heterokaryons in which somatic cell pluripotency gene activation and promoter demethylation occurs rapidly and without cell division. We show that heterokaryon cell fusion based reprogramming is rapid and extensive, and that active DNA demethylation of the somatic OCT4 and NANOG promoters requires Activation Induced Deaminase. Heterokaryon bi-species RNA- Seq identified a role for a secreted factor which enhanced iPS generation and could replace constitutive viral expression of the reprogramming factor c-Myc. These data demonstrate that heterokaryons can serve as a discovery tool for novel reprogramming factors and that findings in cell fusion based reprogramming can inform early events in iPS generation. Mixed species heterokaryons are an attractive model system for investigating mechanisms of active DNA demethylation in the context of the mammalian genome and provide a viable approach for investigating early events in cellular reprogramming.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2013 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Brady, Jennifer Jean |
|---|---|
| Associated with | Stanford University, Department of Microbiology and Immunology. |
| Primary advisor | Blau, Helen M |
| Thesis advisor | Blau, Helen M |
| Thesis advisor | Chen, Chang-Zheng |
| Thesis advisor | Sarnow, P. (Peter) |
| Thesis advisor | Wernig, Marius |
| Advisor | Chen, Chang-Zheng |
| Advisor | Sarnow, P. (Peter) |
| Advisor | Wernig, Marius |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Jennifer Jean Brady. |
|---|---|
| Note | Submitted to the Department of Microbiology and Immunology. |
| Thesis | Ph.D. Stanford University 2013 |
| Location | electronic resource |
Access conditions
- Copyright
- © 2013 by Jennifer Jean Brady
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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