scholarly journals Activation of pancreatic β-cell genes by multiplex epigenetic CRISPR-editing

2020 ◽  
Author(s):  
Gimenez Carla Alejandra ◽  
Curti Lucia ◽  
Hyon Sung Ho ◽  
Grosembacher Luis ◽  
Ross Pablo Juan ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Cellular Differentiation ◽  
Gene Activation ◽  
Directed Differentiation ◽  
Molecular Tools ◽  
Activation Of Transcription ◽  
Simultaneous Activation ◽  
Beta Cell Development ◽  
Per Gene ◽  
Epigenetic Editing

ABSTRACTCRISPR-based systems for epigenetic editing are promising molecular tools that could be harnessed for directed differentiation of pluripotent stem cells. We used the CRISPR/dCas9-VP160, CRISPR/dCas9-TET1 and CRISPR/dCas9-P300 systems for multiplex epigenetic editing and activation of human beta pancreatic genes (PDX1, NEUROG3, PAX4 and INS). The CRISPR/dCas9-P300 system was the most effective at activating genes with reduced number of sgRNA. Using small number of sgRNA per gene was important to induce multiplex gene activation. Combined activation of transcription factors (TFs) involved in beta cell development resulted in INS gene expression; in which sequential TFs activation was more effective than simultaneous activation. Full CRISPR RNA-based delivery system was able to activate all targeted genes. Overall, this study shows the utility of CRISPR tools for epigenetic editing and directed cellular differentiation.

Directed differentiation of mouse-induced pluripotent stem cells generates dopaminergic nerve

2010 ◽  
Vol 34 (8) ◽  
pp. S36-S36
Author(s):  
Ping Duan ◽  
Xuelin Ren ◽  
Wenhai Yan ◽  
Xuefei Han ◽  
Xu Yan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Directed Differentiation ◽  
Induced Pluripotent

scholarly journals Chemically defined and xeno-free culture condition for human extended pluripotent stem cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Bei Liu ◽  
Shi Chen ◽  
Yaxing Xu ◽  
Yulin Lyu ◽  
Jinlin Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Culture Condition ◽  
Human Fibroblast ◽  
Culture System ◽  
Disease Modeling ◽  
Directed Differentiation

AbstractExtended pluripotent stem (EPS) cells have shown great applicative potentials in generating synthetic embryos, directed differentiation and disease modeling. However, the lack of a xeno-free culture condition has significantly limited their applications. Here, we report a chemically defined and xeno-free culture system for culturing and deriving human EPS cells in vitro. Xeno-free human EPS cells can be long-term and genetically stably maintained in vitro, as well as preserve their embryonic and extraembryonic developmental potentials. Furthermore, the xeno-free culturing system also permits efficient derivation of human EPS cells from human fibroblast through reprogramming. Our study could have broad utility in future applications of human EPS cells in biomedicine.

scholarly journals Directed differentiation and long-term maintenance of epicardial cells derived from human pluripotent stem cells under fully defined conditions

2017 ◽  
Vol 12 (9) ◽  
pp. 1890-1900 ◽  
Author(s):  
Xiaoping Bao ◽  
Xiaojun Lian ◽  
Tongcheng Qian ◽  
Vijesh J Bhute ◽  
Tianxiao Han ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Directed Differentiation ◽  
Epicardial Cells ◽  
Term Maintenance

scholarly journals Directed differentiation of human induced pluripotent stem cells into mature kidney podocytes and establishment of a Glomerulus Chip

2018 ◽  
Vol 13 (7) ◽  
pp. 1662-1685 ◽  
Author(s):  
Samira Musah ◽  
Nikolaos Dimitrakakis ◽  
Diogo M. Camacho ◽  
George M. Church ◽  
Donald E. Ingber
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Directed Differentiation ◽  
Induced Pluripotent

scholarly journals Inhibition of DYRK1A disrupts neural lineage specificationin human pluripotent stem cells

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Stephanie F Bellmaine ◽  
Dmitry A Ovchinnikov ◽  
David T Manallack ◽  
Claire E Cuddy ◽  
Andrew G Elefanty ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Gene Dosage ◽  
Gene Activation ◽  
Neural Induction ◽  
Neural Progenitor Cell ◽  
Human Pluripotent Stem Cells ◽  
Mammalian Development ◽  
Neural Lineage ◽  
Early Mammalian Development

Genetic analysis has revealed that the dual specificity protein kinase DYRK1A has multiple roles in the development of the central nervous system. Increased DYRK1A gene dosage, such as occurs in Down syndrome, is known to affect neural progenitor cell differentiation, while haploinsufficiency of DYRK1A is associated with severe microcephaly. Using a set of known and newly synthesized DYRK1A inhibitors, along with CRISPR-mediated gene activation and shRNA knockdown of DYRK1A, we show here that chemical inhibition or genetic knockdown of DYRK1A interferes with neural specification of human pluripotent stem cells, a process equating to the earliest stage of human brain development. Specifically, DYRK1A inhibition insulates the self-renewing subpopulation of human pluripotent stem cells from powerful signals that drive neural induction. Our results suggest a novel mechanism for the disruptive effects of the absence or haploinsufficiency of DYRK1A on early mammalian development, and reveal a requirement for DYRK1A in the acquisition of competence for differentiation in human pluripotent stem cells.

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