title: Generation of T cell receptor-retrogenic mice: improved retroviral-mediated stem cell gene transfer.
abstract: The use of retrogenic mice offers a rapid and flexible approach to T cell receptor (TCR)-transgenic mice. By transducing bone marrow progenitor cells with a retrovirus that encodes a given TCR-α/β subunit, TCR-retrogenic mice can be generated in as few as 4-6 weeks, whereas conventional TCR transgenics can take 6 months or longer. In this updated protocol, we have increased the efficiency of the bone marrow transduction and bone marrow reconstitution compared with our previously published protocol. The main departure from the previous protocol is the implementation of spin transduction with the viral supernatant instead of coculture with the viral producer cell line. The changes in this protocol improve bone marrow viability, increase consistency of the bone marrow transduction and bone marrow engraftment, and they reduce the ratio of bone marrow donor mice to bone marrow recipients.
authors.affiliation: Department of Immunology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
authors.forename: Matthew L
authors.initials: ML
authors.lastname: Bettini
authors.forename: Maria
authors.initials: M
authors.lastname: Bettini
authors.forename: Maki
authors.initials: M
authors.lastname: Nakayama
authors.forename: Clifford S
authors.initials: CS
authors.lastname: Guy
authors.forename: Dario A A
authors.initials: DA
authors.lastname: Vignali
mesh_terms: D000818:Animals; D016026:Bone Marrow Transplantation / Q000379:methods; D018014:Gene Transfer Techniques; D051379:Mice; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics; D013997:Time Factors
chemical_list: D011948:Receptors, Antigen, T-Cell