In fiscal year 2019, 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 F1 hybrid mouse strains, depending on the goal of the experiment. In fall of 2013 the Core started working with the CRISPR/Cas9 technology; it is 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 different guide RNAs into the cytoplasm or pronucleus. The enzyme Cas9, uses the guide RNA to zero in on target DNA, then edits the DNA to disrupt genes or insert a desired sequence. CRISPR/Cas9 technology is a faster, more efficient and a less costly way to make targeted mutation mice. Now, in 2019, approximately 98% of the microinjection performed by the Core is using CRISPR/Cas9 mixes verses the traditional plasmid or BAC constructs. In 2019, 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. In 2019 the TCF started working with the Rare Tumor Initiative. The purpose of this work is to create new mouse models of rare cancers. Creating these mouse models will assist with the studies of rare tumors in hopes of developing new therapies. We will create the mice, maintain the lines, perform the matings, tail biopsies, weanings, inputting the data into a new database and then passing the mice to investigators for testing. In 2019 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. With IVF we can 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. Since August of 2012, our Core has maintained a Cre, FLpe and reporter line 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. We also provide mice to other investigators in CCR & CAPR, as requests come in, always making sure a MTA is filled out if needed. A full list of the services we provide investigators and perform as part of the Core's daily work are included in the following list. We perform microinjection, In vitro fertilization, surgeries (which include embryo transfers, vasectomies, castrations, ovary transfers, ovariectomies, splenectomies, mammary biopsies and mammary fat pad injections), breeding, weaning, tail biopsies, timed pregnancies, necropsies, palpation & measurement of tumors, various types of injections, blood collection, harvesting tissues, 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 also makes its own embryo culture media and anesthetics used within the facility. We wash all our own glassware, make all our own injection needles, holding pipettes and transfer pipettes, we do this to maintain the highest levels of quality control and control costs. 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 which we can take care of just one aspect of a researcher's mouse colony needs or we can manage the entire colony from the creation of the transgenic mouse lines to the lines final disposition and run all their experiments that are covered by the investigator's ASP. In fiscal year 2019 we have performed work for 8 different labs, maintaining experimental mouse lines for three of the 8 labs. The labs we either managed the mouse colonies, trained personnel, set up mouse database programs, run mouse experiments and/or labs we have performed microinjection, in vitro fertilization and/or surgeries for in 2019, are all listed below. Neural Development Section - Lino Tessarollo Hematopoiesis and Stem Cell Biology Section - Jonathan Keller Proteomic Instability of Cancer Section - Chengkai Dai Stem Cell Regulation and Animal Aging Section - Steven Hou CAPR - Serguei Kozlov Regulation of Vertebrate Morphogenesis Section - Susan Mackem Epigenetic Section - Kathrin Muegge Rare Tumor Initiative - Francesco Tomassoni Ardori
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