A transient assay for regulatory gene function in haemopoietic progenitor cells.

2.50
Hdl Handle:
http://hdl.handle.net/10541/90093
Title:
A transient assay for regulatory gene function in haemopoietic progenitor cells.
Authors:
McIvor, Zoe J; Heyworth, Clare M; Johnson, Barbra A; Pearson, Stella; Fiegler, Heike; Hampson, Lynne; Dexter, T Michael; Cross, Michael A
Abstract:
This work aimed to provide a means of assaying directly the effects of transient expression of introduced genes on the survival, proliferation, lineage commitment and differentiation of haemopoietic progenitor cells. For this purpose, we have developed a system that allows isolation of productively transfected, mulitipotent haemopoietic cells within a few hours of the introduction of test genes. We have shown that FDCP-mix cells productively transfected with expression plasmids encoding green fluorescent protein (GFP) differentiate normally and retain colony-forming potential. We constructed an expression vector consisting of a bicistronic cassette in which a GFP marker gene and a test gene are driven from the same promoter. The vector design has been optimized for co-expression and the test gene was shown to be biologically active. The expression profile from a transiently transfected template under different growth conditions reveals that active expression continues for at least 2 d after transfection. The transient transfection of FDCP-mix cells with the vectors described provides a powerful tool for analysis of the immediate early effects of test gene overexpression during haemopoietic differentiation.
Affiliation:
Laboratory of Molecular Medicine, IZKF University of Leipzig, Leipzig, Germany. zoem@medizin.uni-leipzig.de
Citation:
A transient assay for regulatory gene function in haemopoietic progenitor cells. 2000, 110 (3):674-81 Br. J. Haematol.
Journal:
British Journal of Haematology
Issue Date:
Sep-2000
URI:
http://hdl.handle.net/10541/90093
DOI:
10.1046/j.1365-2141.2000.02214.x
PubMed ID:
10997980
Type:
Article
Language:
en
ISSN:
0007-1048
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorMcIvor, Zoe Jen
dc.contributor.authorHeyworth, Clare Men
dc.contributor.authorJohnson, Barbra Aen
dc.contributor.authorPearson, Stellaen
dc.contributor.authorFiegler, Heikeen
dc.contributor.authorHampson, Lynneen
dc.contributor.authorDexter, T Michaelen
dc.contributor.authorCross, Michael Aen
dc.date.accessioned2010-01-19T18:24:34Z-
dc.date.available2010-01-19T18:24:34Z-
dc.date.issued2000-09-
dc.identifier.citationA transient assay for regulatory gene function in haemopoietic progenitor cells. 2000, 110 (3):674-81 Br. J. Haematol.en
dc.identifier.issn0007-1048-
dc.identifier.pmid10997980-
dc.identifier.doi10.1046/j.1365-2141.2000.02214.x-
dc.identifier.urihttp://hdl.handle.net/10541/90093-
dc.description.abstractThis work aimed to provide a means of assaying directly the effects of transient expression of introduced genes on the survival, proliferation, lineage commitment and differentiation of haemopoietic progenitor cells. For this purpose, we have developed a system that allows isolation of productively transfected, mulitipotent haemopoietic cells within a few hours of the introduction of test genes. We have shown that FDCP-mix cells productively transfected with expression plasmids encoding green fluorescent protein (GFP) differentiate normally and retain colony-forming potential. We constructed an expression vector consisting of a bicistronic cassette in which a GFP marker gene and a test gene are driven from the same promoter. The vector design has been optimized for co-expression and the test gene was shown to be biologically active. The expression profile from a transiently transfected template under different growth conditions reveals that active expression continues for at least 2 d after transfection. The transient transfection of FDCP-mix cells with the vectors described provides a powerful tool for analysis of the immediate early effects of test gene overexpression during haemopoietic differentiation.en
dc.language.isoenen
dc.subjectHaematopoietic Stem Cellsen
dc.subject.meshAnimals-
dc.subject.meshCell Culture Techniques-
dc.subject.meshCell Differentiation-
dc.subject.meshCell Line-
dc.subject.meshCell Survival-
dc.subject.meshColony-Forming Units Assay-
dc.subject.meshElectroporation-
dc.subject.meshGene Expression-
dc.subject.meshGenes, Regulator-
dc.subject.meshGenetic Vectors-
dc.subject.meshGreen Fluorescent Proteins-
dc.subject.meshHematopoietic Stem Cells-
dc.subject.meshHumans-
dc.subject.meshLuminescent Proteins-
dc.subject.meshMice-
dc.subject.meshReceptors, Granulocyte-Macrophage Colony-Stimulating Factor-
dc.titleA transient assay for regulatory gene function in haemopoietic progenitor cells.en
dc.typeArticleen
dc.contributor.departmentLaboratory of Molecular Medicine, IZKF University of Leipzig, Leipzig, Germany. zoem@medizin.uni-leipzig.deen
dc.identifier.journalBritish Journal of Haematologyen

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