KI mouse by mouse ES cell genome editing

Mouse ES Cell Genome Editing

  • We generate mouse models with large piece (>2kb) DNA insertion or replacement by editing ES cell genome, then inject modified ES cells into mouse blastocysts for chimeric mice.
  • We use Crispr technology to modify mouse ES cell genome.
  • Available mouse ES cell line genomic background include C57Bl6, 129/ola, and 129fv/fvb.  

User guide:

  1. TKTC performs donor DNA construction, gRNA design, validation, ES cell gene targeting and genotyping.
  2. The positive clones (maximum 4 clones) will be karyotyped and good karyotyped ES cell clones will be injected into mouse blastocysts for chimeric mice. We guarantee germline transmission using TKTC ES cell line.
  3. If user provide gRNAs, that should be validated with >30% cutting efficiency.
  4. If user provides donor DNA, we need 30ug DNA at 1ug/ul concentration. 

ES Cell Microinjection

  • We provide ES cell microinjection into blastocysts service for chimeric mice. 
  • We inject 2-4 positive ES cell clones that are with >60% euploid karyotype.
  • We receive ES cell clones from knockout mouse repositories such as KOMP, EUMMCR and IMPC.
  • User will provide mouse cage card stickers for Sim1 G0421, for pups from ES cell injection.
  • We will update user with ES cell targeting and chimeric mouse info. 
User guide:
  1. For the ES cells that were modified in Stanford TKTC, we will inject 2-4 clones each project into blastocysts and provide at least two high chimeric mice for germline transmission.
  2. For the ES cells from other ES cell repository, user will provide at least one vial of frozen ES cells, that has a good karyotype result and a pathogen free test report. We inject up to 120 blastocysts per service but do not guarantee chimeras or germ line transmission.
  3. For the ES cells from research lab or other facility, we will comply with Stanford Veterinary Service Center guidelines, send the ES cells to VSC for pathogen test prior to blastocyst injection. The testing fee will be charged to the investigator. We inject up to 120 blastocysts per service and do not guarantee chimeras or germ line transmission.
  4. User will provide mouse cage card stickers for Sim1 G0421, pups from ES cell injection will be under user’s APLAC and PTA. 

How to request:

  1. User will provide project goal and set up project strategy together with TKTC.
  2. Log-in iLab, select knock-out/knock-in services, and choose services upon strategy.
  3. Fill in request form, attach project description and related genomic DNA sequences.
  4. Submit request.