title: Retroviral vector production.
abstract: In this unit, the basic protocol generates stable cell lines that produce retroviral vectors that carry selectable markers. Also included are an alternate protocol that applies when the retroviral vector does not carry a selectable marker, and another alternate protocol for rapidly generating retroviral vector preparations by transient transfection. A support protocol describes construction of the retroviral vectors. The methods for generating virus from retroviral vector plasmids rely on the use of packaging cells that synthesize all of the retroviral proteins but do not produce replication-competent virus. Additional protocols detail plasmid transfection, virus titration, assay for replication-competent virus, and histochemical staining to detect transfer of a vector encoding alkaline phosphatase.
authors: Fred Hutchinson Cancer Research Center and Department of Pathology, University of Washington, Seattle, Washington.
authors: A Dusty
authors: AD
authors: Miller
mesh_terms: D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral / Q000235:genetics*; D005819:Genetic Markers; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D051379:Mice; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D019065:Virus Assembly; D014779:Virus Replication
chemical_list: D005819:Genetic Markers
keywords: gene therapy; retroviral vectors; retrovirus packaging cells