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Neurochem Res. 2016 Aug;41(8):2065-74. doi: 10.1007/s11064-016-1918-9. Epub 2016 Apr 25.

Generation of Human Embryonic Stem Cell Line Expressing zsGreen in Cholinergic Neurons Using CRISPR/Cas9 System.

Author information

1
College of Life Sciences, Nanchang University, Nanchang, 330031, China.
2
Stem Cell Translational Research Center, Tongji Hospital, Tongji University School of Medicine, Shanghai, 200065, China.
3
Stem Cell Translational Research Center, Tongji Hospital, Tongji University School of Medicine, Shanghai, 200065, China. siguangli@163.com.
4
College of Life Sciences, Nanchang University, Nanchang, 330031, China. luoyuping@163.com.
5
Stem Cell Translational Research Center, Tongji Hospital, Tongji University School of Medicine, Shanghai, 200065, China. luoyuping@163.com.

Abstract

Lineage specific human embryonic stem cell (hESC) reporter cell line is a versatile tool for biological studies on real time monitoring of differentiation, physiological and biochemical features of special cell types and pathological mechanism of disease. Here we report the generation of ChAT-zsGreen reporter hESC line that express zsGreen under the control of the choline acetyltransferase (ChAT) promoter using CRISPR (Clustered Regularly Interspersed Short Palindromic Repeats)/Cas9 system. We show that the ChAT-zsGreen hESC reporter cell lines retain the features of undifferentiated hESC. After cholinergic neuronal differentiation, cholinergic neurons were clearly labeled with green fluorescence protein (zsGreen). The ChAT-zsGreen reporter hESC lines are invaluable not only for the monitoring cholinergic neuronal differentiation but also for study physiological and biochemical hallmarks of cholinergic neurons.

KEYWORDS:

CRISPR/Cas9 system; Cholinergic neurons; Embryonic stem cell; Reporter cell line; zsGreen

PMID:
27113041
DOI:
10.1007/s11064-016-1918-9
[Indexed for MEDLINE]

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