Dual Notch1 (Variant) -YFP and Inducible Delta-mCherry Reporter CHO-K1 (hN1G4esn) Cell Line

The Notch-Delta signaling pathway plays a vital role in determining cell fates by mediating communication between neighboring cells during development. The Delta ligand has two important roles: it trans-activates Notch in neighboring cells, and cis-inhibits Notch in its own cell. Recent studies suggest that inhibitory same-cell interaction between the Notch receptor and its Delta ligand have significant consequences in the modes of intercellular signaling.The dual reporter cell lines can monitor the transcriptional response of Notch signaling activity owing to its stable Notch receptor expression and corresponding citrine fluorescent protein (YFP) reporter. In addition, the dual reporter cell lines have doxycycline-inducible chimeric Delta (rDII1)-mCherry fusion transmembrane protein, making these cell lines useful as a quantitative platform for analysing gene regulatory circuits in living cells and for better understanding of the network and signaling dynamics in the Notch-Delta system. The hN1 cell line (Cat. No. T3032) stably expresses the human full length Notch1 gene while the hN1G4esn has an intracellular domain of hNotch1 that is replaced with a minimal variant of the transcriptional activator Gal4 to avoid activation of endogenous Notch targets (replacing amino acids 1742 to 2556 of hNotch1 with the amino acids 1-147 and 768-881 of Gal4). Both cell lines show no detectable endogenous Notch or Delta activities and can be induced with 100 ng/ml doxycycline to express Delta-mCherry.