A major focus of research for the next years will be to understand on a more detailed level the molecular mechanisms driving the reprogramming of somatic cells to pluripotent IPS cells. 1. Single-cell analysis of gene expression during cellular reprogramming An unresolved issue is whether activation of specific genes can predict early in the reprogramming process whether a given cell will develop into an iPS cell. Single RNA molecule detection methods will be Used to: a. assess whether a hierarchical program of gene expression leads to IPS cell formation or whether the process is entirely stochastic as suggested by previous observations. b. define markers that at early stages of reprogramming allow the prospective identification of cells that will generate IPS cells. For this, GFP will be inserted into candidate genes to give a marker for prospective isolation of IPS precursors. 2. Stoichiometry of reprogramming factors and quality of iPS cells mRNA mediated reprogramming will be used to systematically titrate the various factors for investigating the effect of factor stoichiometry on the biological properties of the IPS cells. This will allow the optimization of reprogramming with the goal of generating high quality genetically unmodified iPS cells. 3. Transdifferentiation of somatic cells to cells of different lineages Different somatic donor cells such as liver cells and skin keratinocytes will be used for direct conversion into neural precursors and neurons. Stringent reporters will allow the retrospective confirmation of the endodermal arid ectodermal donor cell type.

Public Health Relevance

The disease in the dish approach, based on the IPS technology, is attractive for studying human diseases and for developing novel therapies. However, epigenetic and biological differences between individual IPS cells pose potentially serious hurdles for implementing this approach for research and therapy. This program uses stringent criteria to define the parameters that assure the generation of high quality iPS cells.

National Institute of Health (NIH)
Method to Extend Research in Time (MERIT) Award (R37)
Project #
Application #
Study Section
No Study Section (in-house review) (NSS)
Program Officer
Ravindranath, Neelakanta
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Whitehead Institute for Biomedical Research
United States
Zip Code
Buganim, Yosef; Markoulaki, Styliani; van Wietmarschen, Niek et al. (2014) The developmental potential of iPSCs is greatly influenced by reprogramming factor selection. Cell Stem Cell 15:295-309
Yang, Hui; Wang, Haoyi; Jaenisch, Rudolf (2014) Generating genetically modified mice using CRISPR/Cas-mediated genome engineering. Nat Protoc 9:1956-68
Klemm, Sandy; Semrau, Stefan; Wiebrands, Kay et al. (2014) Transcriptional profiling of cells sorted by RNA abundance. Nat Methods 11:549-51
Armond, Jonathan W; Saha, Krishanu; Rana, Anas A et al. (2014) A stochastic model dissects cell states in biological transition processes. Sci Rep 4:3692
Wang, Haoyi; Yang, Hui; Shivalila, Chikdu S et al. (2013) One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell 153:910-8
Li, Yun; Wang, Haoyi; Muffat, Julien et al. (2013) Global transcriptional and translational repression in human-embryonic-stem-cell-derived Rett syndrome neurons. Cell Stem Cell 13:446-58
Rudenko, Andrii; Dawlaty, Meelad M; Seo, Jinsoo et al. (2013) Tet1 is critical for neuronal activity-regulated gene expression and memory extinction. Neuron 79:1109-22
Wang, Haoyi; Hu, Yueh-Chiang; Markoulaki, Styliani et al. (2013) TALEN-mediated editing of the mouse Y chromosome. Nat Biotechnol 31:530-2
Lodato, Michael A; Ng, Christopher W; Wamstad, Joseph A et al. (2013) SOX2 co-occupies distal enhancer elements with distinct POU factors in ESCs and NPCs to specify cell state. PLoS Genet 9:e1003288
Sarkar, Sovan; Carroll, Bernadette; Buganim, Yosef et al. (2013) Impaired autophagy in the lipid-storage disorder Niemann-Pick type C1 disease. Cell Rep 5:1302-15

Showing the most recent 10 out of 16 publications