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
Transcription
The overall goal of this structure-function method is to dissect at the molecular level the role of different protein domains, or disease-relevance mutations, in naive or differentiated mouse embryonic stem cells. This method can help to reveal ho
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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.