Transgenics and Genetic Constructs Shared Resource

Director: Steve Fiering (Steven.N.Fiering@Dartmouth.edu) 603-653-9966

Director, Cigarette Smoking Exposure Analytical Laboratory: Mardi Crane-Godreau (Mardi Crane-Godreau@Dartmouth.edu) 603-653-9968

Manager: Jen Fields (Jennifer.L.Fields@Dartmouth.edu)

Technician: Sandy Warner (Sandy.Warner@Dartmouth.edu) 603-653-9968

Technical Specialist: Laurie Horne (Laurie.Horne@Dartmouth.edu)

Locations: Director's office: Rubin 622; Animal facility in Borwell Research Building @DHMC

Mission Statement

The mission of the TGCSR is to support the generation and utilization of genetically modified mice by members of the Dartmouth research community.  The support starts with intellectual support in planning and designing transgenic experiments provided by consultation with the TGCSR director.  The TGCSR is now able to offer not only assistance with design of genetic constructs but also the production of genetic constructs for use in transgenic animal production through “recombineering” in either yeast or E. coli.  The tasks of manipulating the mouse embryo are fully provided by the TGCSR either in house or through an established relationship with the Jackson Labs.  Jackson labs is now providing cryopreservation services to protect and preserve valuable transgenic mouse lines.  The TGCSR can also provide genotyping and mouse husbandry services.  As currently organized, the TGCSR can provide the full range of services required to design, generate and maintain transgenic mouse lines. 

Services Provided

• Provide consultation on transgenic mice:
  • construct design and production, overall experimental design,
    utilization, maintenancebreeding and preservation
• Genetic construct generation
  • Use recombineering in E. coli or yeast to generate constructs
    for experimental use
• Oocyte injection transgenic production (transition to Jax)
  • Generate transgenic founders by injection of plasmids, BAC or
    lentivirus into oocytes and transplant the oocytes in foster mothers
• ES cell culture for gene targeting or other purposes
  • Provide a variety of ES cell lines, transfect with KO constructs, select
    clones with drug selection, freeze and provide lysates for genotyping,
    expand and refreeze targeted clones for mouse generation, karyotype
    clones to select best clones
• Generation of chimeric mice from ES cells (transition to JAX)
  • Inject ES cells into blastocysts to generate germline chimeric mice
• Establish lines from frozen embryos from another institutions
  or rederive a pathogen infected transgenic line by embryo transfer
• Genotyping
  • Purify DNA from ES clones (to screen for targeted clones) or tail
    biopsies (to screen for founders or transgenic offspring).
    Establish a genotype assay if not already established.
    Genotype by PCR or Southern blot
• Cigarette Smoke Exposure Analytical Lab
  • Expose rodents to tobacco smoke in the Teague smoke exposure
    facility
• Other services
  • Tail biopsies, tail vein injections, ovariectomies, vasectomies, embryo
    transfer, blood drawing, timed pregnancies, superovulations,
    maintain transgenic colonies (husbandry), maintaining Flp and Cre
    expressing mice for mating to transgenic

Facilities

TGCSR currently maintains two specialized transgenic lines that express Cre or Flp in the germline for genetic manipulation of transgene/knockouts in vivo. TGCSR has a fully equipped embryo manipulation laboratory in the Borwell animal facility and an adjoining 4 room suite of mouse husbandry rooms.