Abstract
Cre-loxp mediated conditional knockout strategy has played critical roles for revealing functions of many genes essential for development, as well as the causal relationships between gene mutations and diseases in the postnatal adult mice. One key factor of this strategy is the availability of mice with tissue- or cell type-specific Cre expression. However, the success of the traditional molecular cloning approach to generate mice with tissue specific Cre expression often depends on luck. Here we provide a better alternative by using bacterial artificial chromosome (BAC)-based recombineering to insert iCreERT2 cDNA at the ATG start of the Upk2 gene. The BAC-based transgenic mice express the inducible Cre specifically in the urothelium as demonstrated by mRNA expression and staining for LacZ expression after crossing with a Rosa26 reporter mouse. Taking into consideration the size of the gene of interest and neighboring genes included in a BAC, this method should be widely applicable for generation of mice with tissue specific gene expression or deletions in a more specific manner than previously reported.
Highlights
Bladder cancer is one of the major causes of malignancy-related morbidity and mortality worldwide, representing the 7th most common type of cancer world wide and the 4th in developed countries [1]
More than 90% of bladder cancers correspond to urothelial carcinomas, since gene mutations are believed to occur in the urothelium, initiating malignant transformation
The causal relationship between these mutations and bladder cancer development has not been well studied in genetic mouse models of bladder cancer due to the lack of suitable transgenic mice that can be used to inactivate these tumor suppressors in the adult, since Pten and Rb are required for embryonic development, and Pten/Rb deficient mice are not born [3,4]
Summary
Bladder cancer is one of the major causes of malignancy-related morbidity and mortality worldwide, representing the 7th most common type of cancer world wide and the 4th in developed countries [1]. The Upk2 promoter has been used successfully in driving expression of Cre [5] and a modified reverse tetracycline transactivator[6] in urothelium specific fashion, as well as in the generation of two transgenic mouse models of invasive and superficial bladder cancers [7,8].
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