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

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
, , , , , , , , , ,
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

Özet

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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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Structure-functionMouse Embryonic Stem CellsRecombinase-mediated Cassette ExchangeRMCEGenome TargetingKnock-outRescue ConstructsCell DifferentiationMoleküler BiyolojiBiotechnology

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Pieters, T., Haenebalcke, L., Bruneel, K., Vandamme, N., Hochepied, T., van Hengel, J., Wirth, D., Berx, G., Haigh, J. J., van Roy, F., Goossens, S. Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange. J. Vis. Exp. (122), e55575, doi:10.3791/55575 (2017).

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