The MGH Transgenic and Gene Targeting Facility generates transgenic and knockout or knock-in mice to study gene function in embryonic stem cell and somatic cell systems. Services include: microinjection of DNA into fertilized embryos for the creation of transgenic mice, transfection of DNA into ES cells for the generation of recombinant ES cell clones, injection of gene-targeted ES cell clones into host blastocysts for the generation of gene knockout or knock-in mice, and other related ES cell based services. The facility is also equipped to perform in vitro fertilization projects, cryopreservation of 8-cell mouse embryos, recovery of cryopreserved mice embryos, and derivation of ES cell lines from a variety of genetic backgrounds. The facility provides consultation within reason on experimental design and vectors for gene targeting-related projects. The facility can also provide advice for the preparation of DNA, recombinant ES clones, mouse genotyping, handling and husbandry. The facility supports innovative and experimental approaches involving transgenic and knockout or knock-in mice to better help investigators study gene regulation and function, and develop appropriate animal models to study human genetic disorders.
Director: Katia Georgopoulos, Ph.D.
Phone: (617) 726-4445
Fax: (617) 726-4453
Associate Director: Lin Wu, Ph.D.
Phone: (617) 724-8124
Fax: (617) 726-4453
Operations Manager: John Seavitt, Ph.D.
Phone: (617) 726-0994 or (617) 724-8280
Fax: (617) 726-4453
Research Technician: Robert Czyzewski
Financial Administrator : Patricia M. Dello Russo
Building 149, Charlestown Navy Yard 13th St. Charlestown, MA
- Microinjection of DNA into fertilized embryos and transplantation of embryos into recipients for the generation of transgenic mice
- Full ES cell transfection service, from DNA transfection into ES cells to picking, establishing, and preparing DNA from the recombinant ES cell clones
- Expansion and storage of established recombinant ES clones
- Microinjection of gene-targeted ES cell clones into blastocysts and transplantation into recipients for the generation of chimeric mice
- Mycoplasma screening of ES cells
- Mouse embryo re-derivation for the generation of pathogen- free mouse lines
- Recovery of cryopreserved mouse embryos
For information on core services and pricing please visit the core's site at http://www.cbrctransgenic.org.
- Contact John Seavitt at 617-726-0994, or 617-724-8280, email@example.com, or Lin Wu at 617-724-8124, firstname.lastname@example.org, to discuss your project, and get a brief orientation of the services.
- Provide a fund number (or PO number for non-partners users) to cover the expenses of your project. Contact Patricia Dello Russo at email@example.com with the payment information and to complete your paperwork.
- Deposit your DNA construct or gene-targeted ES cells and provide information on your project, lab and institution. You will now receive a schedule for your study.
- For transgenic projects: provide a DNA fragment (free of vector) to be injected. It is very important to have a clean (filtered), non-toxic DNA prep with an accurate concentration prior to injection. We need your DNA at a concentration of approximately 50ng/ul in a total volume of approximately 50ul for the injection process. Label your tube with the name of the construct, concentration of the fragment, size of the fragment, and date. We will use a hybrid mouse strain unless you instruct us otherwise.
- For knockout or knock-in projects: provide two tubes of your linearized DNA construct (60 ug DNA in each tube) in 100% ethanol. A picture of the linearized recombinant vector is requested at the time of deposit to ensure quality. A map of the recombinant vector should also be provided to facilitate the experimental procedures (type of selection, method of linearization, and the regulatory elements used to drive selection markers). Label your tube with the name of the construct and date.
- For chimeric mice generation projects: provide two tubes of the frozen gene-targeted ES cells in dry ice or liquid nitrogen, and indicate the background of the ES cells e.g. SV 129 or SV129C57BL/6 hybrid or others. We need to know the name of the ES clone, cell number in each tube to determine seeding surface area per tube, and if any special culture condition is needed for your ES cells.
- The facility will perform the transfection for your knockout or knock-in project, and will notify you as to when to pick up the ES cell DNA samples (~200 colonies per transfection). Your lab is responsible for the screening, and should contact the facility staff with the number of positive clones. Positive clones can then be expanded, DNA generated and returned to you for confirmation. After confirmation please contact the facility to schedule ES cell into blastocyst injection experiments.
- Our facility will contact you after pups (transgenic or gene targeting) are born and inform you of when they are ready to be transferred. You are responsible for the shipping cost. You will contact our animal transfer specialist, and complete the appropriate forms to transfer your mice (within 3 weeks after birth) into your animal facility. If the mice transfer has not occurred after 3 weeks of age, your lab will be charged with additional per-diem rates per cage as determined by our animal facility.