Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from Sox1-gfp knock-in mice.
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.
Aubert
Jerome
J
Institute for Stem Cell Research, University of Edinburgh, King's Buildings, West Mains Road, EH9 3JQ Edinburgh, Scotland.
Stavridis
Marios P
MP
Tweedie
Susan
S
O'Reilly
Michelle
M
Vierlinger
Klemens
K
Li
Meng
M
Ghazal
Peter
P
Pratt
Tom
T
Mason
John O
JO
Roy
Douglas
D
Smith
Austin
A
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D004268:DNA-Binding Proteins / Q000235:genetics*; D005260:Female; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins; D006609:High Mobility Group Proteins / Q000235:genetics*; D017403:In Situ Hybridization; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics; D055748:SOXB1 Transcription Factors; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*
D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D008164:Luminescent Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D055748:SOXB1 Transcription Factors; C098693:Sox1 protein, mouse; D049452:Green Fluorescent Proteins