Cloning and characterization of a retroviral plasmid, pCC1, for combination suicide gene therapy.
Introduction of the herpes simplex virus thymidine kinase (HSV-TK) gene into mammalian cells confers specific sensitivity to killing by the anti-herpes drug ganciclovir (GCV). This gene has therefore been used in a number of cancer gene therapy protocols as a therapeutic gene. However, the therapeutic efficacy of HSV-TK/GCV in cancer gene therapy experiments can be augmented by additional therapeutic genes. We have cloned a retroviral plasmid, pCC1, containing a fusion gene of HSV-TK and Sh-ble driven by an internal simian virus 40 early promoter. This gene encodes a fusion protein that confers GCV sensitivity and Zeocin resistance when introduced into mammalian cells. A multiple cloning site (MCS) allows the introduction of a second therapeutic gene under the transcriptional control of the Moloney murine leukemia virus 5' long terminal repeat. We have generated packaging cell lines electroporated with pCC1 or pCC1 rtIL-2 S (rat interleukin-2 gene cloned in the sense direction in the MCS), the supernatants of which transfer GCV sensitivity only, or both GCV sensitivity and rtIL-2 production, respectively to rat ovarian cancer cells. This plasmid may be useful for the study of combination suicide gene therapy strategies.
Kuiper
M
M
Laboratoire de Thérapie, Génique Humaine et Expérimentale, Centre Jean Perrin, Clermont-Ferrand, France.
Sanches
R
R
Gäken
J A
JA
Bignon
Y J
YJ
D000818:Animals; D003001:Cloning, Molecular; D015774:Ganciclovir / Q000494:pharmacology; D015316:Genetic Therapy; D007376:Interleukin-2 / Q000096:biosynthesis; D010957:Plasmids; D051381:Rats; D012190:Retroviridae / Q000235:genetics*; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000235:genetics
D007376:Interleukin-2; D013937:Thymidine Kinase; D015774:Ganciclovir