Silencing of the somatic cell-type specific genes is a critical yet poorly understood step in reprogramming. To uncover pathways that maintain cell identity, we performed a reprogramming screen using inhibitors of chromatin factors. Here we identify acetyl-lysine competitive inhibitors targeting the bromodomains of coactivators CBP and EP300 as potent enhancers of reprogramming. These inhibitors accelerate reprogramming, are critical during its early stages and, when combined with DOT1L inhibition, enable efficient derivation of human iPSCs with OCT4 and SOX2. In contrast, catalytic inhibition of CBP/EP300 prevents iPSC formation, suggesting distinct functions for different co-activator domains in reprogramming. CBP/EP300 bromodomain inhibition decreases somatic-specific gene expression, histone H3 lysine 27 acetylation (H3K27Ac) and chromatin accessibility at target promoters and enhancers. The master Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:

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Fibroblasts from a Familial Mediterranean Fever (FMF) patient were reprogrammed with episomal vectors by using the Neon Transfection System for the generation of integration-free induced pluripotent stem cells (iPSCs). The resulting iPSC line was characterized to determine the expression of pluripotency markers, proper differentiation into three germ layers, the presence of normal chromosomal structures as well as the lack of genomic integration. A homozygous missense mutation in the MEFV gene (p.Met694Val), which lead to typical FMF phenotype, was shown to be present in the generated iPSC line.

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Induced pluripotent stem cells (iPSCs) offer great promise as tools for basic biomedical research, disease modeling, and drug screening. In this chapter, we describe the generation of patient-specific, transgene-free iPSCs from skin biopsies and peripheral blood mononuclear cells through electroporation of episomal vectors and growth under two different culture conditions. The resulting iPSC lines are characterized with respect to pluripotency marker expression through immunostaining, tested for transgene integration by PCR, and assayed for differentiation capacity via teratoma formation.

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Introduction: Failure and recurrence in breast cancer treatment cause a great obstacle in cancer therapy and identification of cell population named cancer stem cells (CSCs) in the tumor can be led us to define it as target in novel therapeutic strategy. Objectives: The aim of this study is the finding of correlation between stemness and metastatic characteristic, also knowing CSCs as a potential target of therapy because of its developmental behavior and similarities with normal stem cells. Materials and Methods: Here, we focus on the expression of NANOG in breast CSCs, a key molecule in the physiological process of stem cells and the Let-7a that is involved in the differentiation of the cells. Results: In this work, we found that NANOG was highly expressed in SKBR3 and down-regulation of let-7a, as a differentiation miRNA, was found in MDA-MB-468 cells. Conclusion: It will be critical for the developing of effective anti-tumor drugs, utilizing mentioned concepts. Inhibition of NANOG in combination with Let-7a up-regulation can help to decrease the stemness and increase the differentiation of CSCs. The decrease of stemness and increase of differentiation initiate the apoptotic process. So, modification in the mechanism of apoptosis beside anti-cancer drugs provide a good preclinical study goal. However, in order to these drugs become clinical, the problems of their side effects and toxicity must be solved. Differentiation of CSCs provides an optimal condition to activity of immune cells which never let them escape from immune cells by alteration of immunogenicity.