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The gene expression service validates the consequences of the genetic modifications introduced in vivo. Gene expression analyses are specifically pertinent to select transgenic lines established by pronuclear integration (transgene localization, expression level of the transgene, copy number determination, ...), to validate genetically modified mouse models (expected expression of conditional alleles, level of transcripts, splice variants, hypomorphic effects, cell/tissue specific gene inactivation, ...) and to correlate the gene expression analysis with the phenotype of the animals upon clinical evaluation.

Gene expression validation and copy number determination

  • Northern Blot, Southern Blot
  • ddPCR (as replacement of qPCR)

Functional validation

  • Immunohistochemistry
  • Western Blot

Exemple of a fully characterized line: the Insulin1-CreERT2

1. Analysis of Cre and endogenous Ins1 gene expression (qRT-PCR)

Model Validation_gene expression by qRT-PCR.png

2. Endogenous gene expression (triple IHC staining on pancreatic islet)

Model Validation_endogenous gene expression.png

 

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3. Cre activity at the anatomic level (lacZ staining)

Model Validation_Cre activity.png

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4. Clinical phenotyping validation (glucose tolerance test)

Model Validation_Clinical phenotyping.png