Microinjection

Small and large (BAC, YAC) DNA microinjection : production of transgenic rats by microinjection of a DNA construct into a fertilized rat ovocyte of the Sprague- Dawley strain. Approximately 200-250 ovocytes are injected. A minimum of 3 transgenic founders is guaranteed for small DNA.


Microinjection using lentiviral vectors
: Approximately 100-150 ovocytes are injected (required minimal titer 108 UI/ml). A minimum of 3 transgenic founders is guaranteed.

Generation of Knock-out rats using "Zinc-Finger" nucleases or other nucleases.


Genotyping is performed by :

• classical PCR
• Southern-blot.
• Taq-Man (determination of homozygous status)
• genome walking (determination of the insertion site of the transgene within the genome.

Cryopreservation: preservation of lines by freezing of embryos.

Availability of transgenic rats that have already been generated

Technical support (promotors, DNA constructs, screening techniques)

Training and improvement of skills

Development of Procedures

Transfer of technology to industry

Diffusion of information (workshops and congresses)

Immunomonotoring

Cellular immunophenotyping: Cytofluorimetry and/or cryostat immunohistology

Serum concentration of immunoglobulins

In vitro analysis of immune responses

In vivo analyis of immune responses

 

See the flyer for Immunomonitoring of transgenic or KO rats


Principle

 

 

- Construction of DNA, performed by the applicant laboratory
- Advice concerning DNA constructs, performed by the INSERM U643 laboratory
- Purification of the microinjectable DNA
- Superovulation, performed by the TRIP
- Recovery, microinjection and re-implantation of embryos, performed by TRIP
- Tail biopsies of F0 animals, performed by TRIP
- PCR and Southern Blot screening of descendants for the detection of F0 transgenic rats performed by the applicant laboratory
- Differentiation of lines for phenotypic analysis, performed by the applicant laboratory

- Freezing of transgenic embryos, performed by the TRIP
- Hetero/homozygote genotyîng by real-time PCR, performed by TRIP

TRIP is ISO 9001

TRIP is labelised BioGenOuest

 

TRIP is labellised IBISA

Logo IBISA

TRIP is an SFR Bonamy platform

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