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Transgenic/Gene Knockout Shared Resource

The Transgenic/Gene Knockout Facility, located in the Animal Resources Center, employs five full-time technologists who generate genetically engineered mice by pronucleus injection of DNA into fertilized eggs (transgenic mice), by perivitelline space injection of single cell embryos with lentiviral vectors, or by injection of mutated embryonic stem (ES) cells into blastocyst embryos (knockout mice). Investigators supply DNA, lentivirus preparations, or ES cells carrying targeted mutations. The transgenic founder and chimeric mice are bred in the transgenic facility, and offspring are tested in collaboration with the PI for germline transmission of the transgene or the targeted mutation. Genetically modified mice are then transferred to the investigators, who are responsible for further breeding and analysis. The facility also isolates new ES cell lines from any genetically modified mouse line an investigator may have, as well as providing existing ES cell lines suitable for gene targeting experiments, selectable marker plasmids for constructing gene-targeting vectors, and training in ES-cell culture.

Major equipment: The transgenic core facility has four injection rigs, suitable for DNA, lentivirus, and ES cell injections, that consist of a vibration-free table supporting an Olympus inverted microscope outfitted with Narashige micromanipulators and a Narashige pneumatic injector.