MMRRC Testing of the Sanger and Keck MirKO cell lines includes genetic verification at the 3' and/or 5' ends, vector copy number = 1, presence or loss of Y chromosome, and mycoplasma and MPV testing. Based on the average germline rates, we recommend requesting and injecting 2-3 clones per targeted allele. All mutant ES cell lines are heterozygous for the mutation, meaning they carry one modified allele and one wild type allele. MMRRC also recommends performing chromosome counts or karyotyping prior to injection of clones.

Estimated timelines: The repository was sent only one or two vials per clone/cell line from the production center. Processing ES cells (including genetic verification) prior to shipping or injection requires 6-8 weeks. Chromosome counting (to determine % euploid cells) requires an additional 4-8 weeks. Microinjection requires an additional 11-15 weeks. Germline transmission testing (breeding of chimera) requires an additional 8-16 weeks. Germline transmission is not guaranteed.

Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.

The donor or their institution limits the distribution to non-profit institutions only.