Guidelines for Transgenic Mouse Production

Transgenic Mouse Information

The Mouse Genome Engineering core facility asks that all investigators follow the instructions listed below when initiating a new project.

  1. Please provide us with a brief outline of the intended experiment that describes the DNA construct to be injected and the objective or of the experiment. This outline should include a restriction map of your vector and construct.
  2. We request that the investigator must make certain, prior to initiating the transgenic work, that she/he has a DNA screening procedure that will detect a single copy transgene present in mouse genomic DNA. We request this must be demonstrated prior to acceptance and placing the construct into the queue.
  3. We have Institutional Animal Care and Use Committee (IACUC) approval for the necessary manipulations performed by our staff to generate transgenic animals.
  4. You will need to obtain IACUC approval and a protocol number for your intended experiments using your transgenic mice prior to our scheduling your project.
  5. You will also need to obtain an IBC number prior to the scheduling of your project.
  6. MGECF has the expertise to prepare high quality DNA from your plasmid. If you prefer to prepare it yourself,please provide the MGEF with a high quality DNA preparation of your construct, restriction digested to separate all vector sequences. Please provide MGEF with 50μg (in 50-100 μl of pure DDI water) of the DNA construct well labeled with the construct name, concentration and volume and a gel photograph of the digest with the transgene band clearly identified. The facility will also run a test gel of your construct to verify the band and concentration. If you have questions regarding the best procedures to produce the best DNA product for injection, we can provide you with protocols.
  7. We require a cost/center number in order to transfer funds into the MGEF account once we have completed the microinjections.
  8. We will notify you when pups are born and when you should expect to pick up tail biopsies for genotyping. We will clip tails between 8-12 days of age and pups are weaned at 20 days of age. At this time they will be transferred to the investigator. If the genotyping is completed prior to weaning the pups, only the transgene positive mice will be transferred. However, if the genotyping is not completed by weaning age, all of the mouse pups will be transferred to the investigator.
  9. Make arrangements with Comparative Medicine to have space ready for your mice to be transferred. Use the web based RSS Comparative Medicine order form to order a transfer of the mice to your IACUC # and Cost/center and to have the mice transferred to your mouse room. Should you have questions as to how to do this, contact MGECF or Comp Med.

Should you have any further questions please contact CB Gurumurthy. We have protocols that we will provide to you upon request. We have many years of experience with breeding and maintaining genetically altered mouse strains and enjoy sharing our knowledge with you.