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

Xiaoping Bao; Maroof M Adil; Riya Muckom; Joshua A Zimmermann; Aurelie Tran; Natalie Suhy; Yibo Xu; Rocío G Sampayo; Douglas S Clark; David V Schaffer

doi:10.1002/stem.3096

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

Stem Cells. 2019 Dec;37(12):1556-1566. doi: 10.1002/stem.3096. Epub 2019 Nov 13.

Authors

Xiaoping Bao^{1

2

3}, Maroof M Adil^{1

2}, Riya Muckom^{1

2}, Joshua A Zimmermann^{1

2}, Aurelie Tran⁴, Natalie Suhy⁴, Yibo Xu³, Rocío G Sampayo^{1

2}, Douglas S Clark^{2

5}, David V Schaffer^{1

2

3

4}

Affiliations

¹ Department of Bioengineering, University of California, Berkeley, California, USA.
² Department of Chemical and Biomolecular Engineering, University of California, Berkeley, California, USA.
³ Davidson School of Chemical Engineering, Purdue University, West Lafayette, Indiana, USA.
⁴ Department of Molecular and Cell Biology, University of California, Berkeley, California, USA.
⁵ Department of Chemistry, University of California, Berkeley, California, USA.

PMID: 31634414
DOI: 10.1002/stem.3096

Abstract

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.

Keywords: CRISPR/Cas9; Fate mapping; Gene editing; Human pluripotent stem cells; Transcription factors.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems / genetics
Cell Differentiation / genetics
Cell Line
Cell Lineage / genetics*
Gene Editing / methods
Gene Expression Regulation / genetics*
Gene Knock-In Techniques
Gene Targeting
Genes, Reporter / genetics*
Humans
Pluripotent Stem Cells / cytology*
Transcription Factors / metabolism
WT1 Proteins / genetics*

Substances

Transcription Factors
WT1 Proteins
WT1 protein, human