In fiscal year 2015, this Core continues to microinject constructs for the Mouse Cancer Genetics Program and their collaborators; both linear derived fragments of genomic DNA and BAC clones can be microinjected into the pronucleus of inbred and F2 hybrid mouse strains, depending on the goal of the experiment. In Fall of 2013 we started working with the CRISPR/Cas9 technology, the new evolution of creating targeted mutations in mice. With the CRISPR/Cas9 technology mutations can be introduced into a single gene or multiple genes at the same time by injecting gRNAs into the cytoplasm. The enzyme Cas9 uses the guide RNA to home in on target DNA, then edits the DNA to disrupts genes or insert a desired sequence. CRISPR/Cas9 technology is a faster, more efficient and less costly way to make targeted mutation mice. In 2015, and in the years to come, we hope to save the MCGP both time and money with this new CRISPR/Cas9 technology, when it comes to creating targeted mutation mice. We continue to perform in-vitro fertilization (IVF), which we started in fiscal year 2010. IVF of MCGP mouse lines is a way to accelerate colony expansion for experimental research. IVF is a way to rapidly expand a mouse colony to generate large numbers of age, sex & genotypically matched cohorts for experimental use without vastly expanding the number of breeding cages on the shelf. With IVF we can save on cage space, time, and costs as well as accelerate the time it takes to initiate experiments, IVF is basically a more efficient means to expand a mouse colony in a short amount of time. Another way we are trying to save money is to maintain colonies of mice that we use in great quantity or mice that are extremely expensive to buy; those lines include Athymic nudes, Jax B6.CB17-Scid, C57BL/6J, and B6D2F1/J. In August of 2012 our Core started maintaining several Cre and reporter lines used most often by MCGP Labs. With the TFC maintaining these lines there is 1 colony of each line being maintained instead of each lab having their own individual colony and the TCF acts as a central distribution center, (an average cost on many of these Cre or Flpe lines coming from Jax is $232/mouse, just the cost of 4 of these mice would cover the cost of keeping six cages of mice on the shelf for a year. Another line, if retrieved from cryopreservation at Jackson Laboratory, would cost $2525.00/mouse, but to the MCGP investigators it's at no cost because we keep it on the shelf.). We also provide mice to other investigators in CCR & CAPR as requests come in, always making sure MTA's are filled out if needed. We continued performing bleeds for Brad St. Croix and started collecting blood, performing sperm and tissue collection for Jeff Strathern's Lab. We continue to train Post-docs and technicians on various procedures involving the mice and mouse work, as we receive requests. For our microinjection Core, we set up an average of 150 - 200 timed pregnancies per week/40 weeks/year, vasectomize 400 mice per year and send out animal shipments as well as interbuilding transfers, for which we initiated all Material Transfer Agreements for the investigators, as needed. A full list of services we provide investigators include microinjection, IVF, surgeries (which include embryo transfers, vasectomies, castrations, ovary transfers, ovariectomies, splenectomies mammary biopsies, mammary fat pad injections and intraductal mammary gland injections), breeding, weaning, tail biopsies, timed pregnancies, necropsies, palpation & measurement of tumors, injections, blood collection, rederivation, photography, monitoring aging studies, sending animals to histopathology, train technicians, post-docs and investigators how to perform different procedures, initiate material transfer agreements, arrange animal shipments, data entry and database management. The Transgenic Core Facility makes its own embryo culture media and anesthetics used within the facility. We acid wash all of our own glassware, make all our own injection needles, holding pipettes and transfer pipettes. The Transgenic Core Facility also makes all our own microinjection dishes, which allows the injector to work with 150-200 embryos at a time instead of the standard 50 that depression slides hold. The Transgenic Core Facility is truly a multifaceted operation in that we can take care of just one aspect of a researchers mouse colony needs or we can manage the entire colony from the creation of the transgenic mice lines to the lines final disposition and run all their experiments. In 2015 we performed work for 12 different labs, maintaining experimental mouse lines for five of the 12 labs. The labs we either managed the mouse colonies, trained personnel, run mouse experiments and/or labs we have performed microinjection, in vitro fertilization and/or surgeries for in 2015 are all listed below. Neural Development Section - Lino Tessarollo, Tumor Angiogenesis Section - Brad St. Croix, Genetics of Cancer Susceptibility Section - Shyam Sharan, Vascular Biology Section - P. Charles Lin, Cancer Pathways and Mechanisms - Terry Van Dyke, Genetics of Vertebrate Development Section - Mark Lewandoski, Cancer and Developmental Biology Laboratory -Terry Yamaguchi Differentiation and Neoplasia Section - Alan Perantoni Stem Cell Regulation and Animal Aging Section - Steven Hou, Principal Scientist and Program Manager, CAPR - Serguei Koslov, Gene Regulation and Chromosome Biology Laboratory - Jeffery Strathern Genomics and Immunology - Raphael Casellas
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