Insertion of enhanced green fluorescent protein into the lysozyme gene creates mice with green fluorescent granulocytes and macrophages

Pluripotent hematopoietic stem cells have been studied extensively, but the events that occur during their differentiation remain largely uncharted. To develop a system that allows the differentiation of cultured multipotent progenitors by time-lapse fluorescence microscopy, myelomonocytic cells wer...

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Bibliographic Details
Published in:Blood 2000-07, Vol.96 (2), p.719-726
Main Authors: FAUST, N, VARAS, F, KELLY, L. M, HECK, S, GRAF, T
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cell differentiation, maturation, development, hematopoiesis
Cell physiology
Cell Separation
Flow Cytometry
Fluorescence
Fundamental and applied biological sciences. Psychology
Gene Expression
Gene Targeting
Gene Transfer Techniques
Genetic Vectors
Genotype
Granulocytes - cytology
Granulocytes - metabolism
Green Fluorescent Proteins
Hematopoietic Stem Cells - cytology
Luminescent Proteins - analysis
Luminescent Proteins - genetics
Macrophages - cytology
Macrophages - metabolism
Mice
Mice, Transgenic
Molecular and cellular biology
Muramidase - genetics
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Summary:Pluripotent hematopoietic stem cells have been studied extensively, but the events that occur during their differentiation remain largely uncharted. To develop a system that allows the differentiation of cultured multipotent progenitors by time-lapse fluorescence microscopy, myelomonocytic cells were labeled with green fluorescent protein (GFP) in vivo. This was achieved by knocking the enhanced GFP (EGFP) gene into the murine lysozyme M (lys) locus and using a targeting vector, which contains a neomycin resistant (neo) gene flanked by LoxP sites and "splinked" ends, to increase the frequency of homologous recombination. Analysis of the blood and bone marrow of the lys-EGFP mice revealed that most myelomonocytic cells, especially mature neutrophil granulocytes, were fluorescence-positive, while cells from other lineages were not. Removal of the neo gene through breeding of the mice with the Cre-deleter strain led to an increased fluorescence intensity. Mice with an inactivation of both copies of the lys gene developed normally and were fertile. (Blood. 2000;96:719-726)
ISSN:0006-4971
1528-0020
DOI:10.1182/blood.V96.2.719