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

Tim Pieters; Lieven Haenebalcke; Kenneth Bruneel; Niels Vandamme; Tino Hochepied; Jolanda van Hengel; Dagmar Wirth; Geert Berx; Jody J. Haigh; Frans van Roy; Steven Goossens

doi:10.3791/55575

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Biologia dello sviluppo

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

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Biologia dello sviluppo

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JoVE Journal Biologia dello sviluppo

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

10,769 Views

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15:13 min

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April 27, 2017

DOI:

10.3791/55575-v

DOI:

10.3791/55575-v

•

15:13 min

April 27, 2017

•

13 Views

Tim Pieters^1,2,3,4, Lieven Haenebalcke^1,2, Kenneth Bruneel^1,2,4, Niels Vandamme^1,2,4, Tino Hochepied^1,2, Jolanda van Hengel⁵, Dagmar Wirth⁶, Geert Berx^1,2,4, Jody J. Haigh⁷, Frans van Roy^1,2,4, Steven Goossens^1,2,3,4

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

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.

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

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