Mouse development services
Transgenesis Unit
The IMBB transgenesis unit was established in 1995. Transgenic mice are generated via microinjecting linear transgene DNA into the pro nuclei of mouse zygotes. Large DNA fragments are inserted and randomly integrated into the mouse genome. Successful transgenic lines result in the overexpression of the gene of interest.
Gene Targeting Facility
The IMBB Gene Targeting Facility (GTF) offers all services associated with the production through homologous recombination, of conditional or knock-out mice from embryonic stem (ES) cell lines. The facility has generated five projects with germline transmission pups from gene trap and targeted ES cells.
The GTF is located in a separate room within the Animal Cell Culture Facility.
GTF Services
Currently restricted to IMBB users
- Consultation (design of targeting vector, strategy and primers).
- DNA purification.
- Electroporation of ES cells, picking and expanding clones.
- DNA extraction, Southern blot analysis of ES clones.
- Karyotyping.
- Mycoplasma screening.
- Defrosting, expansion of positive ES clones, preparation for blastocyst injection.
- Blastocyst injection of ES clones, chimera production and germline transmission:
Tetraploid complementation: is an alternative method planned for generating ES-cell derived mice using tetraploid cell electro-fusion.
CRISPR/Cas9 technology
Recent advances and improvements in the CRISPR/Cas9-based technology have revolutionized the field of editing the (mouse) genome. Methods such as TAKE and Easi-CRISPR will be used to introduce point mutations, tags, insertions or large deletions using in vitro and in vivo electroporation techniques.
Cryopreservation Unit
Cryopreservation is a method to preserve lines for archiving, shipping, rapidly expanding and an excellent backup method against disease or infertility. The unit has initiated the services of cryopreservation and cryorecovery of sperm from several mouse lines online form. Frozen sperm from these lines will be used for in vitro Fertilization (IVF) followed by vitrification and cryopreservation of the mouse oocytes.