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Struktur-Funktions-Studien in embryonalen Maus-Stammzellen Rekombinase-vermittelten Kassettenaustausch
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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
Proteine enthalten oft mehrere Domänen, die verschiedene zelluläre Funktionen ausüben kann. Gen-Knock-outs (KO) nicht über diese funktionelle Vielfalt berücksichtigen. Hier berichten wir über eine Rekombination vermittelten Kassettenaustausch (RMCE) -basierte Struktur-Funktions-Ansatz in KO embryonalen Stammzellen, die für die molekulare Präparation von verschiedenen funktionellen Domänen oder Varianten eines Proteins ermöglicht.
