High-titer MSCV-based retrovirus generated in the pCL acute virus packaging system confers sustained gene expression in vivo.

Retroviral gene transfer using vectors encoding tumor suppressor genes has been tested repeatedly as a potential anti-tumor therapy. However, most attempts have been hindered by the inability to deliver genes efficiently and to obtain sustained expression in cells growing in vivo. In this paper we describe a method for producing high-titer MSCV virus using the pCL acute retroviral packaging system. This method facilitates the generation of MSCV virus encoding genes that convey the cytostatic or cytocidal phenotypes of benefit in the treatment of cancer. Amphotropic MSCV virus with an average titer of 6 x 10(6) CFU/ml has been routinely produced in this system. We demonstrate that, unlike the pCL retroviral vectors, the MSCV vector is capable of directing sustained in vivo expression of the green fluorescent protein in infected glioma cells following implantation and tumor growth in nude mice.