( Faster easier screening for d...)pFB-ERV: Retroviral Delivery of the Ecdysone Receptor Proteins

Faster, easier screening for difficult transfections when using the complete control kit

Peter Vaillancourt Katherine A. Felts
Stratagene

We describe the vector pFB-ERV, an MMLV-based replication-defective retroviral vector for delivery of the ecdysone receptor proteins RXR and VgEcR. In stable cell lines infected with an estimated single copy of the receptor cassette, induction ratios of greater than 200-fold are attainable in transient reporter transfection assays. The neo-resistance marker is expressed as the third open reading frame (ORF) in a tricistronic CMV expression cassette (the two receptors comprise the first and second ORFs); cell lines harboring single-copy integrants are resistant to as high as 1 mg/ml G418. The CMV promoter ^### is flanked by unique restriction sites and, thus, can be replaced with a cell-type specific promoter of interest.

DNA vector-based systems that allow precise control of gene expression in vivo have become invaluable for the study of gene function in a variety of organisms, particularly when applied to the study of developmental and other biological processes for which the timing or dosage of gene expression is critical to gene function. Such systems have also been successfully used to overexpress toxic or disease-causing genes, to induce gene targeting, and to express antisense RNA. Inducible systems are currently being used by pharmaceutical companies to facilitate screening for inhibitors of clinically relevant biological pathways, and potential applications for gene therapy are being explored.¹

Stratagenes complete control inducible system is based on the insect molting hormone ecdysone, which can stimulate transcriptional activation in mammalian cells harboring the ecdysone receptor protein from Drosophila melanogaster.^2,3 The ecdysone-inducibl
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