Production of avian retroviruses and tissue-specific somatic retroviral gene transfer in vivo using the RCAS/TVA system.
Spatiotemporal retroviral gene transfer into specific somatic mammalian cells using the avian RCAS (replication-competent avian sarcoma-leukosis virus long terminal repeat with splice acceptor)/tumor virus A (TVA) system is a versatile tool for performing lineage tracing and gene function analysis in vivo. RCAS retroviruses carrying the subgroup A envelope transduce only genetically engineered mammalian cells that express the cognate avian retroviral receptor TVA. The RCAS/TVA gene delivery system has been successfully used in various different mouse TVA-expression models. This protocol contains a detailed description of the production of high-titer RCAS retroviruses in chicken fibroblasts and the transduction of proliferating TVA-positive somatic mammalian cells in vivo. By taking advantage of the combination of the RCAS/TVA with the 'universal' Cre/loxP system, the protocol can be used in nearly every proliferating cell type in vivo. The protocol takes 4 weeks from transfection of chicken fibroblasts, which act as the host cells for viral production, to the transduction of TVA-transgenic mice.
Department of Internal Medicine, Technische Universität München, München, Germany.
Alexander
A
von Werder
Barbara
B
Seidler
Roland M
RM
Schmid
Günter
G
Schneider
Dieter
D
Saur
D001355:Alpharetrovirus / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001354:Avian Leukosis Virus / Q000235:genetics; D002645:Chickens; D005347:Fibroblasts / Q000821:virology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008322:Mammals; D051379:Mice; D008822:Mice, Transgenic; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D020079:Terminal Repeat Sequences; D014162:Transfection
D011991:Receptors, Virus