This SRF provides state-of-the-art facilities for the production of transgenic and knockout mice, as well as related rodent embryology techniques. The following procedures are offered on a fee-for-service basis:
Pronuclear injection: Microinjection of purified DNA constructs for the production of transgenic mice. This includes injection of a single transgene construct (plasmid-based or BACs) that has been prepared by the investigator. A minimum of 2 transgenics or 20 live births (or embryos), whichever comes first, is prepared for each set of injections charged to an investigator. Standard transgenic founders are established in either B6C3 hybrids or FVB/N inbreds, while injections can be done in less efficient strains (such as C57Bl/6J) as well.
Gene targeting in ES cells: The Gene Targeting facility has been established to assist investigators with the production of specific targeted loci (i.e., knock-outs and knock-ins) in mouse embryonic stem (ES) cells. The goal of this facility is to electroporate targeting vectors into mouse ES cells to produce clones carrying targeted loci that will be used for the preparation of chimeric mice in the process of establishing novel mouse lines that carry the introduced mutation(s).
ES cell Subcloning: The SRF can use sub-cloning techniques to produce high quality ES cells from lines which have lost their gen line potential. This approach will be useful in cases where ES cells fail to produce geneline transmission due to changes in Karyotype, etc.
ES cell generation from exiting lines: The MGGT SRF can produce ES cells from blastocyst of exiting mouse lines. This is a useful technique for laboratories that would like to generate their own ES cells for in vitro or in vivo studies.
ES cell injection: Microinjection of selected embryonic stem (ES) cell clones into mouse blastocysts to establish chimeric mice for the production of "knockout" mouse lines.
Embryo rederivation: Mouse lines infected with various mouse pathogens can be cleaned up by transfer of embryos to pathogen-free hosts. This will allow these lines to be shipped to collaborators at other institutions or to be moved into cleaner, barrier facilities at Mount Sinai.
IVF rederivation: Mouse lines can be imported into Mount Sinai by obtaining 1-2 males from a collaborator at another institution. These males are used to prepare sperm for in vitro fertilization of wild-type mouse oocytes. The fertilized embryos are transferred to specific pathogen-free hosts to generate clean mice.
Sperm and/or embryo cryopreservation/recovery: The long-term storage of mouse sperm and/or embryos reduces the need for constant maintenance of lines which are not essential for current research needs, as well as providing a backup repository of important lines in case of environmental or weather-related crises.
IVF and frozen embryo recovery: The MGSRF recovers lines from which sperm or embryos have been frozen and stored at Mount Sinai. Cryopreserved sperm samples are maintained in two separate locations to minimize losses in the case of a freezer failure.
Importation of mouse lines through embryos or gametes: The MGSRF has worked successfully with investigators to import lines of mice into Mount Sinai through the use of live, early-stage embryos, frozen embryos or frozen sperm. These methods can offer a relatively quick solution to importation of animals that avoids quarantine issues such quarantine space, and the quarantine time itself.
Information about the SRF's services, protocols and relevant forms can be found here.
To contact the director:
Kevin Kelley, PhD
Tel: (212) 659-6866 or ext 86866