International Knockout Mouse Consortium - Knowledgebase

EUCOMM Gateway protocols High Throughput Gateway (96 Well) Reactions Three Way Gateway Reaction (In tubes)

A number of criteria are applied when choosing the critical exon for a KOMP-CSD or EUCOMM allele, but not every gene can have all criteria fulfilled:

EUCOMM and KOMP-CSD Gene Targeting Cassettes Promoter-Containing Targeting Cassettes L1L2_Bact_P The targeting cassette is flanked by L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate v...

These are the EUCOMM and KOMP-CSD targeting vector plasmid backbones

The clones labelled as “Knockout first” are expected to contain all the loxP sites as shown in the allele map. The knockout is obtained by introduction of a splice acceptor/reporter- cassette with a strong polyA site into an endogenous intron upst...

Question: I am about to generate KO animals using ES cells from KOMP knockout for a particular gene using the KO first allele promoter driven approach. If I am not wrong, I can use these cells directly to generate a KO animal without any treatment...

Summary of all IKMC allele types The wild-type is represented as a transcript with three exons. The ‘floxed’ exon (the exon that will be deleted for the null allele) is exon number 2.

You can find this out yourself by following these steps:

We do a mycoplasma assay on all our clones and one genotype verification using a short-range PCR method. This will take 2-3 more weeks. The parental cell line used to generate the knockout cells has been fully MAP tested, and the actual cells you...

Targeted traps (as we call them) differ from the intended conditional frameshift products in that the LoxP site within the 3’ homology arm has not been confirmed and is likely to be missing (this seems to occur with some regularity with certain pr...

ES Cells

Yes. As part of our quality control effort, EUCOMM does produce 320 different mouse lines. EUCOMM ES cells get selected for mouse production and assigned to four of the five mouse clinics in EUCOMM. Researchers can nominate their favourite genes t...

Yes, the targeting cassette in EUCOMM and KOMP-CSD alleles, is flanked by FRT sites, such that it can be removed independently from the critical exon by means of Flip recombination. This results in the conditional allele (tm1c in the figure). Addi...