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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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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.

Tags

Structure-function Studies Mouse Embryonic Stem Cells Recombinase-mediated Cassette Exchange Protein Domains Disease-relevance Mutations Naive Cells Differentiated Cells Molecular Level Cellular Context Residues Domains Protein Function Rescue Constructs Knock Out Embryonic Stem Cells ES Cells Cell Types Mouse Embryonic Stem Cell Isolation Recombination-based Targeting Factor Assembly Recombination-mediated Cassette Exchange RMCE DNA Fragments Vector Genomic Locus
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