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JoVE Journal Developmental Biology
Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange
 

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

Article DOI: 10.3791/55575-v • 15:13 min • April 27th, 2017
April 27th, 2017 •
1,2,3,4, 1,2, 1,2,4, 1,2,4, 1,2, 5, 6, 1,2,4, 7, 1,2,4, 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

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Summary
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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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Keywords: Structure-function Mouse Embryonic Stem Cells Recombinase-mediated Cassette Exchange RMCE Genome Targeting Knock-out Rescue Constructs Cell Differentiation Molecular Biology Biotechnology
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