Quantitative analysis of gene expression in living adult neural stem cells by gene trapping.
The potential of neural stem cells (NSCs) for the treatment of neurodegenerative diseases makes the identification and characterization of genes involved in neural stem cell responses therapeutically important. Although technologies exist for measuring gene expression in cells, they often provide only a representative expression profile specific to a stimulus and time. We developed a complementary technology based on a retroviral-vector gene-trap approach that uses beta-lactamase-induced disruption of fluorescence resonance energy transfer in the fluorophore CCF-2/AM. A library of 'tagged' adult rat NSCs was generated by transduction with gene-trap virus produced from a single-integrant packaging cell line that allowed us to quantitatively analyze dynamic gene expression changes in real time in living NSCs. Using this library we identified previously unknown genes regulated by oxidative stress, indomethacin and factors that induce differentiation, and show that one of the trapped genes, Sox6, is sufficient to induce astrocytic differentiation when overexpressed.
Scheel
John R
JR
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, California 92037, USA.
Ray
Jasodhara
J
Gage
Fred H
FH
Barlow
Carrolee
C
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D020869:Gene Expression Profiling / Q000379:methods*; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D051381:Rats; D013234:Stem Cells / Q000378:metabolism*