Gene Editing to Generate Versatile Human Pluripotent Stem Cell Reporter Lines for Analysis of Differentiation and Lineage Tracing

Transcription factors (TFs) are potent proteins that control gene expression and can thereby drive cell fate decisions. Fluorescent reporters have been broadly knocked into endogenous TF loci to investigate the biological roles of these factors; however, the sensitivity of such analyses in human plu...

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Bibliographic Details
Published in:Stem cells (Dayton, Ohio) Ohio), 2019-12, Vol.37 (12), p.1556-1566
Main Authors: Bao, Xiaoping, Adil, Maroof M., Muckom, Riya, Zimmermann, Joshua A., Tran, Aurelie, Suhy, Natalie, Xu, Yibo, Sampayo, Rocío G., Clark, Douglas S., Schaffer, David V.
Format: Article
Language:English
Subjects:
Cell differentiation
Cell fate
CRISPR/Cas9
Fate mapping
Fluorescence
Gene editing
Gene expression
Genetic modification
Genome editing
Human pluripotent stem cells
Pluripotency
Progeny
Recombination
Sensitivity analysis
Stem cells
Transcription factors
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Description
Summary:Transcription factors (TFs) are potent proteins that control gene expression and can thereby drive cell fate decisions. Fluorescent reporters have been broadly knocked into endogenous TF loci to investigate the biological roles of these factors; however, the sensitivity of such analyses in human pluripotent stem cells (hPSCs) is often compromised by low TF expression levels and/or reporter silencing. Complementarily, we report an inducible and quantitative reporter platform based on the Cre‐LoxP recombination system that enables robust, quantifiable, and continuous monitoring of live hPSCs and their progeny to investigate the roles of TFs during human development and disease. Stem Cells 2019;37:1556–1566 This study described an efficient two‐step strategy to generate inducible human pluripotent stem cell (hPSC) reporter lines by combining CRISPR/Cas9‐mediated genome editing with the Cre/LoxP system. This iReporter platform enables high‐throughput live cell imaging and fate mapping both in vitro and in vivo.
ISSN:1066-5099
1549-4918
DOI:10.1002/stem.3096