Dual Notch1(Variant)-YFP and Inducible Delta-mCherry Reporter CHO-K1 (hN1G4^(esn)) Cell Line
|T3033||1x106 cells / 1.0 ml|
|Description||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 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.
|Species||Golden Hamster (M. auratus)|
|Species description||Cricetulus griseus (Chinese hamster)|
|Seeding Density||Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.|
For Research Use Only
|Unit quantity||1x106 cells / 1.0 ml|
|Caution||For Research Use Only|
|Cell Type||Drug Discovery Cells|
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10% and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: 37.0°C.
|Preservation Protocol||1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.|
2. Storage Temperature: Liquid nitrogen vapour phase.
1. qRT-PCR was used to assess the expression of Notch mRNA levels in the hN1G4esn and hN1 cells and compared to early T-cell progenitors; 2. The Notch activity induced YFP signal were assess using IgG-Deltaext fusion proteins fixed to plates at different concentrations and time-lapse recording.
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- LeBon, L et al. "Fringe proteins modulate Notch-ligand cis and trans interactions to specify signaling states" Elife Sep 25:3 (2014). PubMed: 25255098 .
- Sprinzak , D et al. "Cis-interactions between Notch and Delta generate mutually exclusive signalling states" Nature 465(7294):86–90 (2010). PubMed: 20418862 .
- Selimkhanov, J et al. "Recent advances in single-cell studies of gene regulation" Curr Opin Biotechnol 23(1):34-40 (2012). PubMed: 22154220.
- Wang, X et al. "Defining single molecular forces required to activate integrin and notch signaling" Science 340(6135):991-4 (2013). PubMed: 23704575.