Cell Stem Cell
Volume 18, Issue 6, 2 June 2016, Pages 755-768
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Article
Genome Editing of Lineage Determinants in Human Pluripotent Stem Cells Reveals Mechanisms of Pancreatic Development and Diabetes

https://doi.org/10.1016/j.stem.2016.03.015Get rights and content
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Highlights

  • Genome editing to study pancreatic lineage determinants in hPSC differentiation

  • A haploinsufficient requirement for PDX1 in pancreatic β cell differentiation

  • Requirements for RFX6 in pancreatic progenitor and endocrine cell differentiation

  • Requirements for NGN3 in the formation of glucose-responsive β cells

Summary

Directed differentiation of human pluripotent stem cells (hPSCs) into somatic counterparts is a valuable tool for studying disease. However, examination of developmental mechanisms in hPSCs remains challenging given complex multi-factorial actions at different stages. Here, we used TALEN and CRISPR/Cas-mediated gene editing and hPSC-directed differentiation for a systematic analysis of the roles of eight pancreatic transcription factors (PDX1, RFX6, PTF1A, GLIS3, MNX1, NGN3, HES1, and ARX). Our analysis not only verified conserved gene requirements between mice and humans but also revealed a number of previously unsuspected developmental mechanisms with implications for type 2 diabetes. These include a role of RFX6 in regulating the number of pancreatic progenitors, a haploinsufficient requirement for PDX1 in pancreatic β cell differentiation, and a potentially divergent role of NGN3 in humans and mice. Our findings support use of systematic genome editing in hPSCs as a strategy for understanding mechanisms underlying congenital disorders.

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Present address: IBEC (Institute for Bioengineering of Catalonia), c/Baldiri Reixac 15-21, 08028 Barcelona, Spain