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

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.

journal_name

Br J Haematol

journal_title

British journal of haematology

authors

McIvor ZJ,Heyworth CM,Johnson BA,Pearson S,Fiegler H,Hampson L,Dexter TM,Cross MA

doi

10.1046/j.1365-2141.2000.02214.x

subject

Has Abstract

pub_date

2000-09-01 00:00:00

pages

674-81

issue

3

eissn

0007-1048

issn

1365-2141

pii

bjh2214

journal_volume

110

pub_type

杂志文章

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