Mouse Model Generation by the Transgenic Facility at the BSRC Alexander Fleming

Transgenic Facility at the BSRC Alexander Fleming

The Transgenic Facility was established in 2001 as the first open-access unit of BSRC Alexander Fleming to provide services in gene modification and the generation of rodent models used in cutting-edge biomedical research. Currently the facility offers services in two areas:

  1. Induction of any type of mutation in mice using specialised instrumentation and protocols for conventional and novel methodologies such as Gene addition, for the production of transgenic lines, Gene targeting or replacement, for the development of new knock-out, conditional knock-out, and knock-in gene models and CRISPR/Cas9 mediated gene editing, for the generation of precision mouse models.
  2. Archiving by Cryopreservation and Distribution, acting as the official national node of EMMA. The Transgenic Facility abides to EMMA principles to offer services in Sperm & Embryo Cryopreservation, Long-term deposition in a state-of-the-art Biobank located at BSRC Alexander Fleming, Cataloguing for Distribution via EMMA or independently, Shipment of Cryopreserved material using the dedicated dry-shipper containers of the Facility, and Revival by In Vitro Fertilisation, Embryo thawing or Embryo Rederivation of live mice.

Service Description:

The Transgenic Facility of BSRC Alexander Fleming provides CRISPR/Cas9 mediated gene editing service for the induction of precision and clinically relevant mutations. The resulted mutations can be indels, exon deletions and point mutation insertions. For the introduction of the specific mutation into the mouse genome, three different sophisticated methods could be used:

  1. Microinjection of the RNPs into the pronucleus or cytoplasm of mouse fertilised oocytes, embryo culture and subsequent embryo transfer.
  2. Embryo electroporation at 1-cell stage embryos, embryo culture and subsequent embryo transfer.
  3. Gonad electroporation (in situ) at first day pregnant females, upon oviduct transfer of the RNPs.

These techniques have been successfully used for the generation of new genome edited mouse models in the framework of Infrafrontier 2020 TA call, as well as upon requests from the scientific community. These precision mouse models are to be used for the study of human diseases such as myotubular myopathy, neurological disorders and various syndromes.

Included in the Service:

Design of sgRNAs and DNA repair templates (if needed) including the prediction of off-target sites, using online CRISPR tools. Performance of microinjections and/or electroporation for the induction of the RNP complexes into mouse embryos. Screening of the resulted animals for the specific mutation, using designed assays (PCR, Sanger sequencing etc). Generation of F0 founder mutant mice. Further mating of founder mice for the provision of at least one F1 genome edited mouse line.

Additional Support

BSRC Alexander Fleming can provide further mating of the F1 mice (usually hybrids) for the back-crossing of the line (e.g. in C57BL6/J or C57BL6/N background).

Relevant Publications
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