Neurochemical Research

, Volume 41, Issue 8, pp 2065–2074 | Cite as

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

  • Jing Zhou
  • Chencheng Wang
  • Kunshan Zhang
  • Yingying Wang
  • Xi Gong
  • Yanlu Wang
  • Siguang LiEmail author
  • Yuping LuoEmail author
Original Paper


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.


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



This work is partially supported by the National Natural Science Foundations of China (Nos. 31571405, 31271450, 31271375 and 31171317) and Jiangxi Natural Science Foundation 20152ACB20008 and 20132BAB204033.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Jing Zhou
    • 1
  • Chencheng Wang
    • 1
  • Kunshan Zhang
    • 2
  • Yingying Wang
    • 2
  • Xi Gong
    • 1
  • Yanlu Wang
    • 1
  • Siguang Li
    • 2
    Email author
  • Yuping Luo
    • 1
    • 2
    Email author
  1. 1.College of Life SciencesNanchang UniversityNanchangChina
  2. 2.Stem Cell Translational Research Center, Tongji HospitalTongji University School of MedicineShanghaiChina

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