@article{38bf6b166d5c4c398f627ccd19d2299d,
title = "Genome Editing of Lineage Determinants in Human Pluripotent Stem Cells Reveals Mechanisms of Pancreatic Development and Diabetes",
abstract = "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.",
author = "Zengrong Zhu and Li, {Qing V.} and Kihyun Lee and Rosen, {Bess P.} and Federico Gonz{\'a}lez and Soh, {Chew Li} and Danwei Huangfu",
note = "Funding Information: We thank Rudolf Jaenisch and Douglas Melton for providing vectors through Addgene; Dirk Hockemeyer for the AAVS1 3′ external probe vector; Alireza Rezania and Sebastian Rieck for advice on differentiating hPSCs into glucose-responsive β-like cells; Daniela Georgieva for technical assistance; Hui Zeng for sharing qPCR primer information; and Shuibing Chen and members of the D.H. laboratory for insightful discussions and critical reading of the manuscript. The MNX1 81.5C10 monoclonal antibody developed by Thomas Jessell and Susan Brenner-Morton was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at the Department of Biology at The University of Iowa. This study was funded in part by the NIH/NIDDK (R01DK096239), New York State Stem Cell Science (NYSTEM C029156), a Basil O{\textquoteright}Connor Starter Scholar Award from March of Dimes Birth Defects Foundation, and the Tri-Institutional Stem Cell Initiative. Z.Z. and F.G. were supported by the NYSTEM postdoctoral fellowship from the Center for Stem Cell Biology of the Sloan Kettering Institute. Publisher Copyright: {\textcopyright} 2016 Elsevier Inc.",
year = "2016",
month = jun,
day = "2",
doi = "10.1016/j.stem.2016.03.015",
language = "English",
volume = "18",
pages = "755--768",
journal = "Cell Stem Cell",
issn = "1934-5909",
publisher = "Cell Press",
number = "6",
}
