[Regulation of gene expression in cultured rabbit corneal stromal cells with recombinant adenovirus using the Cre/loxP system].
OBJECTIVE
To control exogenous gene expression is an important norm in human gene therapy. We attempted to control lac Z gene expression in a rabbit corneal stromal cell (RCS) culture system with recombinant adenovirus (Ad) and the Cre/loxP system.


METHODS
The RCS was transfected with an Ad which expressed the lac Z gene by excisional deletion of stuffer DNA, Ax CA LNLZ. Four days after the transfection, the cells were also transfected with a Cre-producing Ad, Ax CA NCre.


RESULTS
Few cells (0.8%) of the RCS transfected with Ax CA LNLZ expressed the lac Z gene. After the transfection of Ax CA NCre, all cells strongly expressed the lac Z gene. This result showed that recombinant adenovirus using the Cre/loxP system was capable of controlling exogenous gene expression in cultured rabbit corneal stromal cells.


CONCLUSIONS
We suggest that recombinant adenovirus using the Cre/loxP system is useful for studying ophthalmic gene therapy.
Herai
T
T
Department of Bacteriology and Ophthalmology, Iwate Medical University School of Medicine, Morioka, Japan.
Fujiwara
T
T
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D003319:Corneal Stroma / Q000166:cytology*; D005786:Gene Expression Regulation; D019426:Integrases; D007763:Lac Operon; D011817:Rabbits; D011995:Recombination, Genetic; D014162:Transfection; D014764:Viral Proteins
D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases