The Transgenic Core Facility (TCF) provides a centralized resource and state-of-the-art technology for the production of knock-out mice by injection of embryonic stem cells into early mouse embryos, transgenic mice by injection of DNA into the pronucleus of mouse oocytes and also different CRISPR techniques.
Laser assisted ES-Cell Injection:
- Culture of the cells until they are ready for injection
- ES-Cell Laser Injection into 8-cell wildtype mouse embryo stages
- Embryo transfer into the oviduct of pseudo-pregnant recipients
Pronuclear Injection:
- Production of cycle-timed Zygotes by using superovulation in your needed mouse background
- Microinjection of Plasmids or BAC’s into the pronuclear of fertilized Zygotes
- Embryo transfer into the oviduct of pseudo-pregnant recipients
CRISPR Technologies:
- CRISPR electroporation into murine ES-Cells
- CRISPR into Zygotes (cytoplasm or pronuclear)
- CRISPR electroporation in Zygotes
- iGONAD (improved genome-editing via oviductal nucleic acids delivery)
Rederivation/Import of mouse lines:
- Cleaning of infected mouse lines by embryo transfer
- Preferred import of frozen embryos
- We also find solutions for hygienically critical imports
In Vitro Fertilisation:
- Rescue of mouse line with bad breeders
- Import of mouse lines via frozen sperm samples
- Fast track from STR analyzed chimeric sperms to germline offsprings
Backup-Freezing:
- Sperm freezing (10 straws per mouse line, incl. an IVOS protocol (sperm analyser)
- Embryo freezing (150 until 250 embryos in different straws)
ES-Cell Derivation:
- Derivation of pluripotent ES-Cell lines from Blastocyst stages
- 3-5 different cell lines, incl. sex determination
- Deriving of MEF’s (Mouse embryonic Fibroblasts)
ES-Cell Electroporation:
- Homologues recombination by Electroporation of DNA constructs
- Selection, splitting and freezing of two master-plates
- In-house Genotyping
Glass Capillary Studio:
- Pulling, grinding, bending and fire-polishing or spiking of Glass needles
- Using filamented and non-filamented Borosilicate-Glass
