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Regenerative Medicine (2003) / Chapter Skim
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5 Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from Sox1-gfp knock-in mice
Pages 20-25

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From page 20...
... Institute for Stem Cell Research, University of Edinburgh, King's Buildings, West Mains Road, EH9 3JQ Edinburgh, Scotland; tThe Scottish Centre for Genomic Technology and Informatics, University of Edinburgh, Chancellor's Building, 49 Little France Crescent, EH16 4SB Edinburoh, Scotland; and §Division of Biomedical Sciences, University of Edinburgh, Hugh Robson Building, George Square, EH8 9XD Edinburgh, Scotland The transcription factor Sox1 is the earliest and most specific known marker for mammalian neural progenitors. During fetal development, Sox1 is expressed by proliferating progenitor cells throughout the central nervous system and in no tissue but the lens.
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From page 21...
... Microarray and Sequence Analysis. A subtracted library enriched for genes expressed during retinoic acid-induced neural commitment of ES cells (7)
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From page 22...
... brain; BM, bone marrow; CC, coiled coil; CRD, cysteine-rich domain; Dev, developing; DRG, dorsal root ganglia; E, embryo; Ep, epithelial; HLH, helix-loop-helix; HMG, high-mobility group; KH, K homology RNA-binding domain; LRR, leucine-rich repeat; M, muscle; Mes, mesenchymal; NB, Northern blot; N PCs, neural precursor cells; NS, nervous system; NT, neural tube; OV, optic vesicle; PN, postnatal; RRM, RNA recognition motif; S skeletal; TM, transmembrane; ZnF, zinc finger.
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From page 23...
... Together, these results confirm the efficient separation of neural and nonneural cell populations by flow cytometry. P311 and Riken cDNA2810027019 represent two genes that are induced in retinoic acid treated embryoid bodies (74.
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From page 24...
... We then carried out a pilot microarray screen with the aim of identifying genes specifically expressed both during neural commitment of ES cells and in neural progenitor cells in vivo. From 384 arrayed SSH clones, we identified 15 unique clones showing preferential expression in the GFP-positive cell population.
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From page 25...
... Previously described expression profiles and our in situ hybridization data of the differentially regulated clones examined confirms the underlying principle of using RNA prepared from Soxl-selected cells to screen custom-built microarrays enriched for neural genes. This pilot scale screen has been sufficient to highlight several genes, notably Nhlh2, Lrrnl, Hrmtll3, Rtnl, and the unknown gene corresponding to the P2H5 EST, for further investigation as potential regulators of neural development.
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Key Terms

  • situ hybridization
  • cell populations
  • embryoid bodies
  • neural precursor
  • marker genes

  • neural precursors
  • stem cells
  • microarray screen
  • preferentially expressed
  • adult neural

  • precursor cells
  • neural stem
  • progenitor cells
  • subgranular layer
  • embryonic stem

  • targeting vector
  • neural differentiation
  • neural progenitors
  • flow cytometry
  • correctly targeted

  • dentate gyrus
  • neural tissues
  • cell sorting
  • stem cell
  • retinoic acid

  • cell population
  • root ganglia
  • cells throughout
  • dorsal root
  • subtracted library


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