S'identifier

JoVE Journal

Cancer Research

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Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells

2.5K Views

09:16 min

May 5th, 2022

May 5th, 2022

2,500 Views

0:04

Introduction

0:45

Dissection of Mouse Bladder

2:09

Dissociation of Urothelial Cells

4:41

Counting and Resuspending Cells

5:27

Adenoviral Transduction and Plating Organoids

7:06

Results: Characterization of Triple Knockout Organoids via Ex Vivo Adenovirus-Cre Recombinase

8:31

Conclusion

Transcription

We developed a rapid and efficient protocol to generate bladder cancer organoids via ex vivo gene editing of normal mouse urothelial cells carrying floxed alleles in genes of interest. This ex vivo method has a one-to-two-week workflow instead of

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This protocol describes the process of the generation and characterization of mouse urothelial organoids harboring deletions in genes of interest. The methods include harvesting mouse urothelial cells, ex vivo transduction with adenovirus driving Cre expression with a CMV promoter, and in vitro as well as in vivo characterization.

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