In Vivo and In Vitro Tissue‐Specific Expression of Green Fluorescent Protein Using the Cre‐Lox System in Mouse Embryonic Stem Cells

Embryonic stem cells (ES) are pluripotent and may therefore serve as a source for the generation of specific cell types required for future therapies based on cell replacement. The isolation of defined cell populations from a certain lineage or tissue is a prerequisite for the analysis of the potential of such ES‐derived cells in animal transplantation studies. Here, using the Cre/loxP system, we report the generation of murine ES cells conditionally expressing the hrGFP gene at the cell surface. Such ES cells can be differentiated in vitro into neurons displaying GFP activity in neurites. Transgenic mice derived from these ES cells permit the targeting of GFP‐expression to specific tissues and provide material from the three germ layers suitable for molecular and biochemical analysis.

[1]  T. Shimazaki,et al.  Isolation and transplantation of dopaminergic neurons generated from mouse embryonic stem cells , 2004, Neuroscience Letters.

[2]  M. Stoffel,et al.  Navigating the pathway from embryonic stem cells to beta cells. , 2004, Seminars in cell & developmental biology.

[3]  A. Colman Making new beta cells from stem cells. , 2004, Seminars in cell & developmental biology.

[4]  M. Li,et al.  Generation of embryonic stem cells and transgenic mice expressing green fluorescence protein in midbrain dopaminergic neurons , 2004, The European journal of neuroscience.

[5]  S. Chiba,et al.  Transplantation of Motoneuron-Enriched Neural Cells Derived from Mouse Embryonic Stem Cells Improves Motor Function of Hemiplegic Mice , 2003, Cell transplantation.

[6]  H. Schöler,et al.  Derivation of Oocytes from Mouse Embryonic Stem Cells , 2003, Science.

[7]  Marius Wernig,et al.  Tau EGFP embryonic stem cells: An efficient tool for neuronal lineage selection and transplantation , 2002, Journal of neuroscience research.

[8]  H. Wichterle,et al.  Directed Differentiation of Embryonic Stem Cells into Motor Neurons , 2002, Cell.

[9]  R. McKay,et al.  Dopamine neurons derived from embryonic stem cells function in an animal model of Parkinson's disease , 2002, Nature.

[10]  Kazuto Kobayashi,et al.  Dynamics of tyrosine hydroxylase promoter activity during midbrain dopaminergic neuron development , 2002, Journal of neurochemistry.

[11]  Bruce G. Jenkins,et al.  Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[12]  Caiying Guo,et al.  A cre recombinase transgene with mosaic, widespread tamoxifen‐inducible action , 2002, Genesis.

[13]  Hideyuki Okano,et al.  Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[14]  P. Chambon,et al.  An efficient system for conditional gene expression in embryonic stem cells and in their in vitro and in vivo differentiated derivatives , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[15]  P. Gruss,et al.  Pax4 regulatory elements mediate beta cell specific expression in the pancreas , 2001, Mechanisms of Development.

[16]  M. Capecchi,et al.  Fgf8 is required for outgrowth and patterning of the limbs , 2000, Nature Genetics.

[17]  K. Mizuseki,et al.  Induction of Midbrain Dopaminergic Neurons from ES Cells by Stromal Cell–Derived Inducing Activity , 2000, Neuron.

[18]  J. Miyazaki,et al.  A novel reporter mouse strain that expresses enhanced green fluorescent protein upon Cre‐mediated recombination , 2000, FEBS letters.

[19]  A. McMahon,et al.  Modification of gene activity in mouse embryos in utero by a tamoxifen-inducible form of Cre recombinase , 1998, Current Biology.

[20]  P. Gruss,et al.  The Pax4 gene is essential for differentiation of insulin-producing β cells in the mammalian pancreas , 1997, Nature.

[21]  S. Sugano,et al.  Efficient gene activation in mammalian cells by using recombinant adenovirus expressing site-specific Cre recombinase. , 1995, Nucleic acids research.

[22]  R Kemler,et al.  The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. , 1985, Journal of embryology and experimental morphology.

[23]  A. Bradley,et al.  Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines , 1984, Nature.

[24]  M. Kaufman,et al.  Establishment in culture of pluripotential cells from mouse embryos , 1981, Nature.

[25]  A. Mansouri Determination of gene function by homologous recombination using embryonic stem cells and knockout mice. , 2001, Methods in molecular biology.

[26]  B. Sauer,et al.  Conditional gene knockout using Cre recombinase. , 2001, Molecular biotechnology.

[27]  J. B. Houseknecht,et al.  Analysis of Cell-Cycle Profiles in Transfected Cells Using a Membrane-Targeted GFP , 1999 .

[28]  S. Brenner,et al.  Distinct cis-essential modules direct the time-space pattern of the Pax6 gene activity. , 1999, Developmental biology.

[29]  K. Rajewsky,et al.  A cre-transgenic mouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells. , 1995, Nucleic acids research.