The state-of-the-art centralized MGH Gene Targeting Facility established in 1995 generates knockout mice for studies on gene regulation and function in embryonic and adult development. The transfection service provides the technology and expertise necessary for the preparation of recombinant ES Clones and their subsequent injection into host blastocysts. The facility also serves as a major educational resource for investigators on training on new techniques, vectors and experimental design. The facility will continue to design innovative and experimental approaches involving knockout mice to better help investigators study gene function and develop appropriate animal models to study human genetic disease. The Core Director, Dr. Katia Georgopoulos, and her staff are available for consultation.

Director: Katia Georgopoulos, Ph.D.
Phone: (617) 726-4445
Fax: (617) 726-4453
Email: katia.georgopoulos@cbrc2.mgh.harvard.edu

Associate Director: Lin Wu, Ph.D.
Phone: (617) 724-8124
Fax: (617) 726-4453
Email: lin.wu@cbrc2.mgh.harvard.edu

Other Staff:

Operation Manager: Jeffrey Wu
Phone: ( 617) 726-0994
Email: jwu0@partners.org

Major Equipment:

• Gene pulsor electroporation machine
• Incubators
• Centrifuges
• Hoods
• Microinjection apparatus

• Full ES Cell transfection service from DNA transfection to DNA preparation
• Microinjection of positive ES Cell clones
• Derivation of ES Cell lines from mutant mice
• 129 phage genomic library
• Consultation on targeting vector construction

1. Contact Dr. Katia Georgopoulos at katia.georgopoulos@cbrc2.mgh.harvard.edu or at
   (617) 726-4445 or Jeffrey Wu at (617) 726-0994 who will discuss your project and provide a brief orientation of the facility and services.
2. Contact Patty Dello Russo at (617) 726-4431 to complete paperwork and agreement.
3. Contact Jeffrey Wu at (617) 726-0994 to arrange deposit of construct and arrange timing of the transfection.
4. Drop off DNA sample: 2 tubes containing 60 ug each (ie. 120 ug total), suspended in 100% ethanol. A picture of the digested DNA vector is requested at the time of deposit to review quality. Information (i.e. MAP) on the nature of the vector should also be communicated to facilitate the experimental procedures (type of selection, method of linearization, and the regulatory elements used to derive it) as well as grant numbers.
5. The core facility will perform the transfection and will notify you as to when to pick up the ES cell DNA samples (50-200 colonies per transfection).
6. Your lab is responsible for the screening and should contact the ES Cell Core Staff on the number of positive clones. The generated DNA will be returned to you for further confirmation.
7. After your lab has verified the identity of the ES Cell clone, the facility will schedule the blastocyst injection.
8. Your lab will be contacted once chimeric mice are born.
9. Appropriate forms will be completed by you and the facility and your mice will be transferred back to you 3 weeks after their birth. If mice have not been transferred within 3 weeks of birth, your mice will be charged appropriate additional per-diem rates per cage as charged by the animal facility.