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Derivation of Haploid Neural Stem Cell Lines by Selection for a Pax6-GFP Reporter.
Journal article

Derivation of Haploid Neural Stem Cell Lines by Selection for a Pax6-GFP Reporter.

  • Gao Q 1 State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University , Tianjin, China .
  • Zhang W 1 State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University , Tianjin, China .
  • Ma L 1 State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University , Tianjin, China .
  • Li X 1 State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University , Tianjin, China .
  • Wang H 1 State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University , Tianjin, China .
  • Li Y 1 State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University , Tianjin, China .
  • Freimann R 3 Institute of Molecular Health Sciences, Swiss Federal Institute of Technology Zurich , Zurich, Switzerland .
  • Yu Y 2 Reproductive Medical Center, Department of Gynecology and Obstetrics, Peking University Third Hospital , Beijing, China .
  • Shuai L 1 State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University , Tianjin, China .
  • Wutz A 3 Institute of Molecular Health Sciences, Swiss Federal Institute of Technology Zurich , Zurich, Switzerland .
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  • 2018-02-24
Published in:
  • Stem cells and development. - 2018
CRISPR
Pax6-GFP
haploid
neural stem cells
nontoxic sorting
Animals
Cell Differentiation
Cells, Cultured
Female
Gene Expression
Genes, Reporter
Green Fluorescent Proteins
Haploidy
Karyotype
Mice
Mice, 129 Strain
Mouse Embryonic Stem Cells
Nestin
Neural Stem Cells
PAX6 Transcription Factor
English Haploid cells facilitate genetic screening of recessive mutations for a single set of chromosomes. Haploid embryonic stem cells (haESCs) have been achieved in several species and widely utilized in genetic screens. The fact that haESCs undergo substantial diploidization during differentiation has limited the screening to other haploid cell types. In this study, we report a method to establish haploid neural stem cells (haNSCs) by selection for a Pax6 reporter. We inserted a green fluorescence protein (GFP) marker gene by homologous recombination into the Pax6 locus of an haESC line. GFP-positive haploid cells could be sorted and further cultured in the NSC medium for more than 30 passages. The established haNSCs expressed neural lineage markers and could differentiate into neurons, oligodendroglia, and astrocytes. Our study shows the feasibility of deriving haploid proliferative somatic cell lines using a genetically encoded reporter that suggest a system for genetic screening of neural and retinal development.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://folia.unifr.ch/global/documents/112923
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