Services Provided:
1. Generation of Transgenic Founders from DNA Constructs. Details of methodology may be found elsewhere (Hogan B, Beddington R, Constantini F, Lacy E. Manipulation of the Mouse Embryo. A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, 1994). DNA transgenic constructs, supplied by the investigator, will be microinjected into zygotes. Tails from potentially transgenic offspring will be analyzed by the investigator to determine whether transgenic founders have been produced. If necessary, repeat injections of the construct will be performed to ensure that the investigator is supplied with at least 3 founder animals. The founder animals, their breeding and their phenotypic analysis will become the responsibility of the investigator. The cost of the DNA microinjection service will be $250 per construct for FVB mice and $1,750 per construct for C57Bl/6 mice. Please inquire about prices for other strains.
2. Cryopreservation of Mutant Mice. From time to time investigators may wish to cryopreserve embryos of valuable transgenic lines. The investigator will supply the Transgenic Core with 4 genetically modified male mice. These mice will be mated with superovulated females (of the same genetic background) and embryos will be harvested and frozen at the 8-cell stage utilizing a slow rate freezer and DMSO as the cryoprotectant. The cryopreserved embryos will be held in straws containing a maximum of 20 embryos per straw. A minimum of 600 embryos will be frozen per transgenic line. After freezing to -35 degrees centigrade, the straws will be transferred to canes and stored in liquid nitrogen cryofreezers. The embryos will be stored within the transgenic facility (current capacity is 120,000 embryos). Following freezing, 100 of the 600 embryos will be thawed and transferred to pseudopregnant recipients to assess for viability. Currently, survivability of frozen embryos in the BIDMC Transgenic Facility is ~ 90%. The cost of the freezing service will be $250 per line.
3. Microinjection of Targeted ES cell clones. The BADERC investigator will provide the transgenic facility with targeted ES cell clones. These targeted clones will be microinjected into 3.5 day old mouse embryos (blastocysts). The DERC investigator will then receive, in return, chimeric mice derived from these ES cell clones, which will then be used, by the DERC investigator, to create heterozygous and homozygous gene knockout mice. The cost of the ES cell microinjection service will be $250 per injection of two ES cell clones.
