Genetic Engineering and Model Validation

Our mission is to develop the most adapted, highly physiologically relevant animal models for your research program. The genetic engineering services are flexible and extensive to help you generate your custom-made RAT or MOUSE model, from the design of vectors construction to the functional validation of the model. Additional services are proposed for genotyping. Our deliverables are transgenic or chimera animals, in SPF or SOPF status.

Our services involve:

  • Design of an optimized strategy including prediction of off-target sites
  • Preparation of all materials for injection (DNA, vectors, sgRNA, Cas9 mRNA, ES cells...)
  • Injection of genetic materials to generate F0 mutated animals
  • Genotyping of F0 animals and delivery of F0 candidates (CRISPR/Cas9)
  • Germline transmitted F1 animals
  • Off-target analysis in F0 founder animals (CRISPR/Cas9, optional)
  • Germline breeding of F0 founders to achieve germ line transmission (CRSIPR/Cas9, optional)
  • Off-target analysis in germline transmitted F1 animals (CRISPR/Cas9, optional)

 

Genetic Engineering150 vectors per year

  • Knock-out constitutive, conditional
  • Knock-out tissues specific and/or inducible
  • Knock-in, genetic targeted insertion :
    Point mutation, complex and multi-allelic modifications, reporter Tag (Rosa26, HPRT, Random transgenesis,...), depletion of cell types (hDTR, DTa chains,...), interactoms, cell tracking.
  • Humanization
  • Nuclease genome engineering (CRISPR/Cas9)

All mouse strains and lines provided by PHENOMIN are available on this page.

ES Cells service200 mutant ES clones and 250 ES validations per year

  • Derivation of WT and mutant new ES cell lines from different genetic background (C57BL/6N, Sv129/PAS and BALB/cN ES)
  • In vitro differentiation pathway
  • Mutant ES clones production
  • ES validation
  • In vitro analysis of gene function
  • Extensive quality controls to secure your mouse model and injection of EUCOMM / KOMP / IKMC mutant clones

R&D program: ES cell differentiation, rat ES cells

We provide you with expert advices, we define a targeted strategy in accordance with your needs and ensure a high scientific and technical output.

Microinjection450 projects per year

  • Injection of DNA into fertilized eggs from random insertion of transgenes
  • Injection of validated ES cell into blastocysts
  • Efficient germ line transmission rate

Genotyping service60 000 samples per year

  • Fully automated genotyping plateform
  • Validation of genetic modification
  • Confirmation of mutation by sequencing
  • Short range PCR or qPCR 
  • Easy-to-use genotyping protocol provided for each lines generated

Validation of genetic modified mouse model

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 Functional validation
Northern Blot, Southern Blot Immunohistochemistry
ddPCR (as replacement of qPCR) Western Blot