Construction of a safe and efficient retrovirus packaging cell line

Ecotropic and amphotropic retrovirus packaging cell lines have been constructed in which the helper virus genome have been separated onto two plasmids, and the psi packaging signal and 3' LTR have been removed. The gag and pol genes on one plasmid and the env gene on another plasmid were transfected into NIH 3T3 cells. Packaging cell lines produced by these transfected genes released titers of replication-defective retroviral vectors which were comparable to titers produced by packaging cell lines containing the helper virus genome on one plasmid. There has been no evidence of recombination events between the ecotropic helper virus plasmids and the vector virus plasmid that would result in the generation of intact replication-competent virus. These results suggest that a packaging cell line containing gag, pol and env on different plasmids is efficient and safe for use in retroviral gene gransfer.