InfrafrontierGR/Phenotypos

The InfrafrontierGR-Phenotypos project supports eligible customers with a free-of-charge mouse model development service implemented as a Trans-regional Access activity supporting a total of two (2) projects in this call.The access unit offered covers the production of a minimum of a single heterozygous mouse line, carrying the gene alteration of choice. The model development services will be run by the BSRC Fleming Transgenics Facility (2 access units) and may involve one of the three main mouse transgenic technologies:
- CRISPR/Cas9 method for the production of genome-edited mouse line. The service includes prediction of off-target sites, preparation of sgRNA’s and Cas9 mRNA/protein, and delivery into zygotes to generate F0 founder mutant animals. Selected F0 animals will be bred to germ line to produce F1 genome edited animals. Possible allele types that can be generated should be small insertions or deletions (indels).
- Blastocysts injection method of gene-targeted embryonic stem cells (ESC) for the generation of a new mouse line carrying a disrupted or mutated genetic locus. Already validated gene-targeted ES cell clones must be provided from either international consortia such as IKMC, or other sources, at the Investigator’s cost. For each project a minimum of two independent ES cell clones will be requested to maximize the probability to obtain a germline-transmitting chimera. Breeding of chimeras result in the generation of the F1 mutated founder line.
- DNA microinjection method in the pronucleus of zygotes for the generation of transgenic mice, carrying an inserted cassette in their genome. The DNA construct will be provided by the investigator accompanied by a proof that the transgene can be detected in genomic DNA by a functional assay, as well as a gel photo where integrity and quantity of the DNA is documented with the help of a known DNA marker. The minimum of 3 transgenic founders will be delivered for every construct injected.

Newly developed mouse models will be made available to selected applicants within an average of 12 months following provision of all required information to start the mouse production. The generated mouse models will be made available to the scientific community. An optional grace period of up to 1 year for mouse resources may apply, with immediate release of mouse resources after expiry of the grace period. Mouse mutant lines will be deposited into the INFRAFRONTIER/EMMA repository for subsequent use by the scientific community.

A collaboration agreement will be established between BSRC Fleming and applicants.