Stable reconstitution of IKK-deficient mouse embryonic fibroblasts.
Retroviral transduction is an invaluable technique in molecular biology used to express proteins encoded by nonviral genes in mammalian cells. A key feature of this technique is the ability to create cell lines that stably express the protein of interest and can be cultured long term. Here we describe a retroviral transduction procedure for mouse embryonic fibroblasts (MEFs) that uses Platinum-E cells to rapidly package high-titer, helper-free retrovirus. This technique is useful to study the role of key signaling kinases in the NF-κB signal transduction pathway.
Department of Animal Biology, The University of Pennsylvania School of Veterinary Medicine, 3800 Spruce Street OVH200E, Philadelphia, PA, 19104, USA.
Carolyn M
CM
Gray
Michael J
MJ
May
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D004789:Enzyme Activation; D005347:Fibroblasts / Q000378:metabolism*; D015870:Gene Expression; D055786:Gene Knockout Techniques; D005822:Genetic Vectors / Q000235:genetics; D051550:I-kappa B Kinase / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D014162:Transfection / Q000379:methods
D011993:Recombinant Fusion Proteins; D051550:I-kappa B Kinase