C-terminal deletion of NOTCH1 intracellular domain (N1^ICD) increases its stability but does not amplify and...

Since the generation of a mouse strain conditionally expressing the active intracellular domain of Notch1 (N1^ICD), many laboratories have exploited this model (Rosa^N1-ICD) to assess the impact of constitutive Notch1 signalling activation in normal and pathological processes. It should be underscored that Cre-recombination leads to the expression of a C-terminally truncated form of N1^ICD (N1^ICDdC) in the Rosa^N1-ICD mutant mice. Given that no studies were undertaken to delineate whether deletion of this region leaves intact N1^ICD function, stable cell lines with single targeted integration of inducible N1^ICD and N1^ICDdC were generated. We found that C-terminal deletion of N1^ICD stabilized the protein but did not promote the activity of Notch responsive promoters. Furthermore, despite higher expression levels, N1^ICDdC failed to phenocopy N1^ICD in the promotion of anchorage-independent growth. Our results thus suggest that the C-terminal region of N1^ICD plays a role in shaping the Notch response. Therefore, it should be taken into consideration that N1^ICD is truncated when interpreting phenotypes of Rosa^N1-ICD mutant mice.