Retroviral delivery of the ecdysone-regulatable gene expression system.
We describe a set of Moloney Murine Leukemia Virus (MoMLV)-based replication-defective retroviral vectors for delivery of the ecdysone-inducible system into mammalian cells. The vector pFB-ERV contains a tricistronic CMV expression cassette from which the ecdysone receptor proteins RXR and VgEcR are expressed, with the neo-resistance marker expressed as the third open reading frame (ORF). The inducible vector pCFB-EGSH contains an ecdysone-inducible expression cassette inserted between the viral LTRs in the antisense orientation relative to that for the viral promoter. Potential interference from the proviral 5' LTR is obviated due to a SIN deletion in the 3' LTR. When used together, induction ratios of over 1000-fold were achieved in NIH3T3 cells using firefly luciferase as a reporter.
Vaillancourt
Peter
P
Stratagene Cloning Systems, 11011 N. Torrey Pines Rd., La Jolla, California 92037, USA. pvaillancourt@AMEvolution.com
Felts
Katherine A
KA
D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004440:Ecdysone / Q000096:biosynthesis* / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D007668:Kidney / Q000196:embryology / Q000378:metabolism*; D008979:Moloney murine leukemia virus / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*
D011994:Recombinant Proteins; D004440:Ecdysone