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Generating Neural Stem Cells from iPSCs with Dopaminergic Neurons Reporter Gene

  • Hyenjong Hong
  • Marcel M. DaadiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1919)

Abstract

Genetic reporters offer attractive approaches to investigate gene expression, gene function, and spatiotemporal assessment in vitro and in vivo. Tyrosine hydroxylase (TH) is the rate-limiting enzyme for the biosynthesis of the dopamine neurotransmitter, and thus it has been used as a reliable marker for dopaminergic neurons in vitro and in vivo. Herein we describe a method for making iPSC lines with TH-green fluorescent protein reporter gene using CRISPR/Cas9 technique.

Key words

Tyrosine hydroxylase Reporter CRISPR/Cas9 Human pluripotent stem cells Neural stem cell Parkinson’s disease 

Notes

Acknowledgment

The authors thank members of the Daadi laboratory for the helpful support and suggestions. This work was supported by the Worth Family Fund, the Perry & Ruby Stevens Charitable Foundation and the Robert J., Jr. and Helen C. Kleberg Foundation, the NIH primate center base grant (Office of Research Infrastructure Programs/OD P51 OD011133), and the National Center for Advancing Translational Sciences, National Institutes of Health, through Grant UL1 TR001120.

Disclosures: Dr. Marcel M. Daadi is founder of the biotech company NeoNeuron.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Southwest National Primate Research CenterTexas Biomedical Research InstituteSan AntonioUSA
  2. 2.Department of Radiology, Research Imaging Institute, Cell Systems and Anatomy, Long School of MedicineUniversity of Texas Health Science Center at San AntonioSan AntonioUSA

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