A new packaging cell line for SV40 vectors that eliminates the generation of T-antigen-positive, replication-competent recombinants.
Simian virus 40 (SV40) vectors are efficient vehicles for gene delivery to hematopoietic and hepatic cells. To ensure their replication incompetence and because of safety considerations, it is critical that the vectors do not contain T-antigen sequences. Available packaging cell lines for T-antigen replacement vectors, COS and CMT4, contain considerable sequence identity with the vectors, leading to homologous recombination and reacquisition of the T-antigen gene. We constructed a packaging cell line, COT18, with minimal sequence identity to the vector. Vector stocks produced by passaging on COT18 had high transducing activity and undetectable levels of T-antigen-positive, replication-competent contaminants. This cell line provides a means for the preparation of safe SV40 vector stocks.
Arad
Uri
U
Department of Hematology, Hebrew University-Hadassah Medical School, Hadassah University Hospital, Jerusalem, Israel.
Ben-Nun-Shaul
Orly
O
El-Latif
Mahmoud Abd
MA
Nissim
Orna
O
Oppenheim
Ariella
A
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000502:physiology*; D019556:COS Cells; D002522:Chlorocebus aethiops; D005822:Genetic Vectors / Q000235:genetics*; D011995:Recombination, Genetic; D013539:Simian virus 40 / Q000235:genetics* / Q000502:physiology; D014161:Transduction, Genetic; D014709:Vero Cells; D014779:Virus Replication
D000952:Antigens, Polyomavirus Transforming