Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from Sox1-gfp knock-in mice

Jerome Aubert, Marios P. Stavridis, Susan Tweedie, Michelle O'Reilly, Klemens Vierlinger, Meng Li, Peter Ghazal, Tom Pratt, John O. Mason, Douglas Roy, Austin Smith

    Research output: Contribution to journalArticle

    186 Citations (Scopus)

    Abstract

    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. We generated a reporter mouse line in which egfp is inserted into the Sox1 locus. Sox1GFP animals faithfully recapitulate the expression of the endogenous gene. We have used the GFP reporter to purify neuroepithelial cells by fluorescence-activated cell sorting from embryonic day 10.5 embryos. RNAs prepared from Sox1GFP+ and Sox1GFP- embryo cells were then used to perform a pilot screen of subtracted cDNAs prepared from differentiating embryonic stem cells and arrayed on a glass chip. Fifteen unique differentially expressed genes were identified, all previously associated with fetal or adult neural tissue. Whole mount in situ hybridization against two genes of previously unknown embryonic expression, Lrrn1 and Musashi2, confirmed the selectivity of this screen for early neuroectodermal markers.
    Original languageEnglish
    Pages (from-to)11836-11841
    Number of pages6
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume100
    Issue numberSuppl. 1
    DOIs
    Publication statusPublished - 2003

    Fingerprint

    Embryonic Structures
    Fluorescence
    Neuroepithelial Cells
    Embryonic Stem Cells
    Fetal Development
    Lenses
    Genes
    In Situ Hybridization
    Glass
    Flow Cytometry
    Transcription Factors
    Stem Cells
    Central Nervous System
    Complementary DNA
    RNA
    Gene Expression

    Cite this

    Aubert, Jerome ; Stavridis, Marios P. ; Tweedie, Susan ; O'Reilly, Michelle ; Vierlinger, Klemens ; Li, Meng ; Ghazal, Peter ; Pratt, Tom ; Mason, John O. ; Roy, Douglas ; Smith, Austin. / Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from Sox1-gfp knock-in mice. In: Proceedings of the National Academy of Sciences of the United States of America. 2003 ; Vol. 100 , No. Suppl. 1. pp. 11836-11841.
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    abstract = "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. We generated a reporter mouse line in which egfp is inserted into the Sox1 locus. Sox1GFP animals faithfully recapitulate the expression of the endogenous gene. We have used the GFP reporter to purify neuroepithelial cells by fluorescence-activated cell sorting from embryonic day 10.5 embryos. RNAs prepared from Sox1GFP+ and Sox1GFP- embryo cells were then used to perform a pilot screen of subtracted cDNAs prepared from differentiating embryonic stem cells and arrayed on a glass chip. Fifteen unique differentially expressed genes were identified, all previously associated with fetal or adult neural tissue. Whole mount in situ hybridization against two genes of previously unknown embryonic expression, Lrrn1 and Musashi2, confirmed the selectivity of this screen for early neuroectodermal markers.",
    author = "Jerome Aubert and Stavridis, {Marios P.} and Susan Tweedie and Michelle O'Reilly and Klemens Vierlinger and Meng Li and Peter Ghazal and Tom Pratt and Mason, {John O.} and Douglas Roy and Austin Smith",
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    Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from Sox1-gfp knock-in mice. / Aubert, Jerome; Stavridis, Marios P.; Tweedie, Susan; O'Reilly, Michelle; Vierlinger, Klemens; Li, Meng; Ghazal, Peter; Pratt, Tom; Mason, John O.; Roy, Douglas; Smith, Austin.

    In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100 , No. Suppl. 1, 2003, p. 11836-11841.

    Research output: Contribution to journalArticle

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    AU - Aubert, Jerome

    AU - Stavridis, Marios P.

    AU - Tweedie, Susan

    AU - O'Reilly, Michelle

    AU - Vierlinger, Klemens

    AU - Li, Meng

    AU - Ghazal, Peter

    AU - Pratt, Tom

    AU - Mason, John O.

    AU - Roy, Douglas

    AU - Smith, Austin

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    AB - 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. We generated a reporter mouse line in which egfp is inserted into the Sox1 locus. Sox1GFP animals faithfully recapitulate the expression of the endogenous gene. We have used the GFP reporter to purify neuroepithelial cells by fluorescence-activated cell sorting from embryonic day 10.5 embryos. RNAs prepared from Sox1GFP+ and Sox1GFP- embryo cells were then used to perform a pilot screen of subtracted cDNAs prepared from differentiating embryonic stem cells and arrayed on a glass chip. Fifteen unique differentially expressed genes were identified, all previously associated with fetal or adult neural tissue. Whole mount in situ hybridization against two genes of previously unknown embryonic expression, Lrrn1 and Musashi2, confirmed the selectivity of this screen for early neuroectodermal markers.

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