Dlx5 induces expression of COL1A1 promoter contained in a retrovirus vector.
OBJECTIVE
To determine whether retrovirally expressed Dlx5, a homeobox-containing transcription factor, can induce a 2.3 kb rat COL1A1 promoter-reporter construct, which is transduced into osteoblastic cells by the use of a retrovirus vector.


METHODS
A self-inactivating retrovirus vector containing the rat COL1A1 driving green fluorescent protein (GFP) was transduced into chick calvarial periosteal cells. These cells were then infected with a replication-competent retroviral vector expressing Dlx5, or a control vector. The cells were cultured in the presence of ascorbic acid and beta-glycerol-phosphate, which promotes osteoblastic differentiation. Expression of the COL1A1 promoter was assessed by detecting GFP with fluorescence microscopy.


RESULTS
GFP was detected only in cells infected with the Dlx5 expressing retrovirus. The GFP positive cells were observed in regions of the culture that had undergone osteoblastic differentiation, as detected by cell morphology and the presence of a mineralized matrix.


CONCLUSIONS
The 2.3 kb rat COL1A1 promoter fragment contains elements responsive to Dlx5, and the self-inactivating retroviral vector allows these elements to be used appropriately.
Tadić
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Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington CT 06030, USA.
Erceg
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Stover
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Rowe
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Lichtler
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D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D002642:Chick Embryo; D003094:Collagen / Q000378:metabolism*; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D018398:Homeodomain Proteins / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D051381:Rats; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection
D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; C501120:Dlx5 protein, mouse; D018398:Homeodomain Proteins; D003094:Collagen