title: COS-1 cells as packaging host for production of lentiviruses.
abstract: We present a protocol for in vitro production of recombinant lentiviruses using COS-1 African green monkey kidney epithelial cells and HEK293T human embryonic kidney epithelial cells as packaging cells. COS-1 and HEK293T express SV40 large T antigen, amplifying transfected circular plasmids harboring SV40 replication origin. Support protocols for evaluation of transfection efficiency by in situ β-galactosidase enzyme activity assay and titer of infection-capable virions are also provided. Advantages of using COS-1 packaging cells over the standard HEK293T cells for contamination-sensitive applications or automated processing are discussed.
authors.affiliation: Massachusetts General Hospital Center for Cancer Research, Charlestown, Massachusetts, USA.
authors.forename: Crystal J
authors.initials: CJ
authors.lastname: MacKenzie
authors.forename: Toshi
authors.initials: T
authors.lastname: Shioda
mesh_terms: D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D016086:Lentivirus / Q000235:genetics*; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation / Q000379:methods*
chemical_list: D000952:Antigens, Polyomavirus Transforming