Endogenous RAC1 P29S in mouse melanocytes

We studied the effects of long-term endogenous expression of the melanoma oncogene RAC1 P29S using melanocyte cultures isolated from transgenic mice Overall design: Transgenic mice with a conditional knock-in of the P29S mutation in the endogenous Rac1 locus were generated and crossed onto C57BL/6J, Tyr-CreER;Trp53fl/fl mice. Melanocytes were isolated from the dorsal skin of neonatal mice and cultured using the protocol developed by others (Godwin et al. Current Protocols in Cell Biology, 1.8.1-1.8.20, 2014). Melanocytes were recombined by 4OH-tamoxifen treatment. Three independent melanocyte cultures from Tyr-CreER+/-;Trp53fl/fl;Rac1LSL-P29S/wt mice were used and compared to two independent cultures from Tyr-CreER+/-;Trp53fl/fl;Rac1wt/wt mice. The spontaneously immortalised C57BL/6J melanocyte cell line melan-a was used as the third independent replicate for the Rac1 wt/wt group.