Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange

10.9K Views

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15:13 min

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April 27th, 2017

DOI :

10.3791/55575-v

April 27th, 2017

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Tim Pieters¹^,²^,³^,⁴, Lieven Haenebalcke¹^,², Kenneth Bruneel¹^,²^,⁴, Niels Vandamme¹^,²^,⁴, Tino Hochepied¹^,², Jolanda van Hengel⁵, Dagmar Wirth⁶, Geert Berx¹^,²^,⁴, Jody J. Haigh⁷, Frans van Roy¹^,²^,⁴, Steven Goossens¹^,²^,³^,⁴

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

Summary

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Keywords Structure function

Mouse Embryonic Stem Cells

Recombinase mediated Cassette Exchange

Chapters in this video

0:05

Title

1:11

The RMCE Principle

2:15

Isolation of RMCE-compatible Knockout Mouse ES Cells

5:22

Isolate RMCE-compatible Knockout ES Cells

7:29

Generation of an RMCE-compatible Targeting Vector Using Recombination Cloning

9:25

RMCE-mediated ES Cell Targeting of Rescue Constructs to the R26 Locus and Differentiation of ES Cells in Embryoid Bodies

12:41

Results: Cystic EB Formation Using Recombinase Mediated Cassette Exchange

14:27

Conclusion

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