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Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange
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1Department of Biomedical Molecular Biology,Ghent University, 2Inflammation Research Center,VIB, 3Center for Medical Genetics,Ghent University Hospital, 4Cancer Research Institute Ghent (CRIG), 5Department of Basic Medical Sciences, Faculty of Medicine and Health Sciences,Ghent University, 6Helmholtz Center for Infection Research, 7Mammalian Functional Genetics Laboratory, Division of Blood Cancers, Australian Centre for Blood Diseases, Department of Clinical Haematology,Monash University and Alfred Health Alfred Centre
Chapters
- 00:05Title
- 01:11The RMCE Principle
- 02:15Isolation of RMCE-compatible Knockout Mouse ES Cells
- 05:22Isolate RMCE-compatible Knockout ES Cells
- 07:29Generation of an RMCE-compatible Targeting Vector Using Recombination Cloning
- 09:25RMCE-mediated ES Cell Targeting of Rescue Constructs to the R26 Locus and Differentiation of ES Cells in Embryoid Bodies
- 12:41Results: Cystic EB Formation Using Recombinase Mediated Cassette Exchange
- 14:27Conclusion
Proteins often contain multiple domains that can exert different cellular functions. Gene knock-outs (KO) do not consider this functional diversity. Here, we report a recombination-mediated cassette exchange (RMCE)-based structure-function approach in KO embryonic stem cells that allows for the molecular dissection of various functional domains or variants of a protein.
