Transgenic Core Facility provides services for generating transgenic and knockout mice. Established in 1992, the BWH Core Facility has been providing gene targeting and microinjection services using state-of-the-art facilities and equipment.

Director: Arlene Sharpe, M.D, Ph.D.
Phone: (617) 432-6569
Fax: (617) 432-6570
Email: asharpe@rics.bwh.harvard.edu

Other Staff:

Transgenic Specialist: Lina Du, M.D.
Phone: (617) 432-6227
Email: lina_du@hms.harvard.edu

ES Cell Culture Technician: Xiaomeng Tang
Phone: (617) 432-6182
Email: xtang@rics.bwh.harvard.edu

ES Cell Culture Technician: Matthew Finn
Phone: (617) 432-6576
Email: matthew_finn@hms.harvard.edu

Major Equipment:

The Core Facility has all of the equipment necessary for generation of transgenic mice including a Nikon Diaphot microscope equipped with Nomarski Optics and Narishige micromanipulators for microinjections, Nikon surgical microscopes for egg isolation ad transfer, a sutter needle puller and a de Fonbrune microforge.

• Microinjection of DNA constructs (including BACs) into pronuclei for generating transgenic mice (FVB or C57 BL/6 strain background)
• Microinjection of embryonic stem (ES) cells into blastocysts for generating knockout mice
• ES cell transfection for generating ES cell clones with the targeted combination event
• Lentiviral transgenics
• Microinjection of IPS
  

Our ES facility will generate targeted ES cell lines with KO constructs provided by investigators. Upon receipt of the constructs and application form, electroporation will be scheduled. The Core Director, Dr. Arlene Sharpe will prioritize the service requests according to the difficulty of the project and work load.

1. Transfection of your linearized DNA construct into ES cells, J1 (129/SV) and Bruce 4 (C57 BL/6J).
2. After electroporation and selection, clones will be picked, cultured in multi-well plates and replicated. The clones will be frozen to be used to expand positive clones.
3. The investigator will be notified and given DNA isolated from from each clone suitable for screening.
4. Since storage of frozen cells in multi-well dishes is not optimal, it is strongly suggested that researcher return the screening results to the core within 4 months of receipt of the DNA samples.
5. After screening the clones, up to six ES positive clones will be expanded and frozen down.
6. After expansion, the investigator will then receive DNA from each clone for reconfirmation of targeted recombination. After reconfirmation, the best growing clones will then be scheduled for injection into blastocysts.  Chimeric mice will be given to the investigator.

Please complete the appropriate application forms below for your service request:

Application for Injection of DNA into Pronuclei

Injection of Embryonic Stem Cells into Blastocysts

In vitro Gene Targeting Application

Preparation of DNA construct for microinjection

To access antibody request form, please click on link below:

http://transgenic.partners.org