Generation, cryopreservation and rederivation of transgenic mouse lines

This application covers training and performing of generation, cryopreservation and rederivation of genetically modified mouse lines. Currently we send our mouse injection constructs to University of Oregon (Eugene, OR, USA) and generate our transgenic mouse lines there, then send them to Norway where we are maintaining the lines at our transgenic facility in active breeding. We wish to be able to do the mouse transgenesis and line preservation / rederivation experiments here, so that we can greatly reduce the cost and stress of animals from long distance shipping and the number of animals being kept in overlapping colonies in two countries. In addition, being able to do cryopreservation and rederivation of mouse lines will give us the possibility to freeze all mouse lines that are not currently in use and revive them whenever needed. This will cut down the number of animals used for line maintenance, and ensure the quality of our transgenic lines by limiting genetic drift and therefore limit the effects of drift on our experimental reproducibility and research quality. Currently we have around 60 unique transgenic mouse lines, to cryopreserve all of them, we will need about 3000 mice over a 4-year period. Animals involved in survival surgeries will be treated under anesthesia during the surgeries and receive analgesia before and after the surgeries. Therefore, moderate distress is expected. All other procedures involve minimal distress. Training and supervision of our practicing will be provided by highly experienced specialist Ute Hostick, who is Manager and Injector of Transgenic Mouse Facility (TMF) at the University of Oregon (UO). Ute has been working with mice more than 20 years, have produced over one hundred unique transgenic mouse lines and performed countless rederivations as well as cryopreservation of embryos/sperm since she started working at the TMF in 2003.