Generation of Transgenic Mice

Generation of Transgenic Mice

The Transgenic Core Facility at the Penn State College of Medicine (PSCOM) provides a centralized facility for the production of transgenic mice. DNA microinjection produces random insertion transgenic mice. The Transgenic Core will perform all the steps necessary to produce transgenic mice, including set-up, embryo harvesting, DNA microinjection and transfer to recipient mothers. The investigator is responsible for supplying the purified construct (DNA) prepared using our DNA Guidelines and DNA Preparation Protocol.

We will complete one set of injections (minimum of 150 viable embryos implanted into recipient females, 20 candidate pups, or 2 founder pups, whichever criteria is met first) and supply the investigator with tail biopsies of the resulting pups. The investigator is responsible for genotyping potential founder mice. Once founder mice have been identified, the Transgenic Core will transfer the animals to the investigator.

The standard mouse strains used for microinjection are B6D2F1, FVB/N and C57Bl/6J or the investigator can provide the mice with no guarantee. If the investigator requires microinjections on another background strain, this can be arranged for an additional cost. For additional information please refer to the Transgenic Core Facility Guidelines.

For fee schedule, see Transgenic and Knockout Mouse Generation Fees.

Statistics

Blastocyst injections: 1997-present

  • 24 cell lines producing chimeras
  • 125 chimeras from 989 pups born- 12.6% success rate
  • At least 10 lines have been reported to be germline

Transgenic injections

2005-present

  • 20 injections
  • 188 pups tested
  • 29 positive founders; 15.4% success rate

(Before requiring DNA quality Check) 1997-2004

  • 299 injections
  • 2242 pups tested
  • 224 positive founders-10.0% success rate

The Transgenic Core Facility has injected 83 constructs for 23 investigators, generating 253 transgenic founders, averaging 3 founders per construct.

Rederivations: 2005-present

  • 13 rederivations for 6 investigators
  • 112 pups born-8.6 pups per rederivation
  • 56 positive pups-4.3 positive pups per rederivation

Major Equipment

  • Leica Labovert FS microscope with dual micromanipulators
  • Sutter Flaming/Brown Micropipette Puller P-87
  • Narashige microforge MF-9
  • Olympus SZH10 dissecting stereoscope
  • Eppendorf Transinjector 5246
  • Laminar flow hood
  • VWR CO2 incubator Model 2310

Related Guidelines & Protocols

  • Transgenic Core Facility Guidelines
  • DNA Guidelines
  • DNA Preparation Protocol