In vivo analysis of cell polarity during cell-cell fusion

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Among various phosphatidylinositols (PI) on the plasma membrane, the most abundant product, PI(4,5)P2 and locally-produced PI(3,4,5)P3 and PI(3,4)P2 seem to be particularly important in polarity formation of the cells. The PH domain of PLC delta1 binds to PI(4,5)P2 while the PH domain of Akt binds to PI(3,4)P2 and PI(3,4,5)P3, which is locally produced depending on PI3-kinase activity. By using these domains, I succeeded in observing the polarity of fusing cells. To apply this system in vivo, KH2 ES cells which can express those domains fused to GFP or RFP in response to doxycyclin were generated. Now that genetic quality of the ES cells were confirmed, the mice who express the polarity probe is being generated.