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Transgenic Res. 2018 Feb;27(1):75-85. doi: 10.1007/s11248-018-0058-1. Epub 2018 Jan 30.

Generation of a biotinylatable Sox2 mouse model to identify Sox2 complexes in vivo.

Author information

1
Department of Pediatric Surgery, Erasmus Medical Center-Sophia Children's Hospital, Wytemaweg 80, 3015 CN, Rotterdam, The Netherlands.
2
Netherlands Proteomics Center, Erasmus Medical Center, Rotterdam, The Netherlands.
3
Department of Pediatric Surgery, Erasmus Medical Center-Sophia Children's Hospital, Wytemaweg 80, 3015 CN, Rotterdam, The Netherlands. r.rottier@erasmusmc.nl.
4
Department of Cell Biology, Erasmus Medical Center, Rotterdam, The Netherlands. r.rottier@erasmusmc.nl.

Abstract

Sox2 is a Sry-box containing family member of related transcription factors sharing homology in their DNA binding domain. Sox2 is important during different stages of development, and previously we showed that Sox2 plays an important role in branching morphogenesis and epithelial cell differentiation in lung development. The transcriptional activity of Sox2 depends on its interaction with other proteins, leading to 'complex-specific' DNA binding and transcriptional regulation. In this study, we generated a mouse model containing a biotinylatable-tag targeted at the translational start site of the endogenous Sox2 gene (bioSox2). This tag was biotinylated by the bacterial birA protein and the resulting bioSox2 protein was used to identify associating partners of Sox2 at different phases of lung development in vivo (the Sox2 interactome). Homozygous bioSox2 mice are viable and fertile irrespective of the biotinylation of the bio tag, indicating that the bioSox2 gene is normally expressed and the protein is functional in all tissues. This suggests that partners of Sox2 are most likely able to associate with the bioSox2 protein. BioSox2 complexes were isolated with high affinity using streptavidin beads and analysed by MALDI-ToF mass spectrometry analysis. Several of the identified binding partners are already shown to have a respiratory phenotype. Two of these partners, Wdr5 and Tcf3, were validated to confirm their association in Sox2 complexes. This bioSox2 mouse model will be a valuable tool for isolating in vivo Sox2 complexes from different tissues.

KEYWORDS:

Biotinylatable tag; In vivo protein complexes; Knock-in; Sox2

PMID:
29383478
PMCID:
PMC5847153
DOI:
10.1007/s11248-018-0058-1
[Indexed for MEDLINE]
Free PMC Article

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