Transgenic Core

Director:

Bradford B. Lowell, MD, PhD Professor of Medicine, BIDMC and Harvard Med. School
E-mail: blowell@bidmc.harvard.edu

Co-Director:

Dong Kong, PhD Instructor, BIDMC and Harvard Med. School
E-mail: dkong@bidmc.harvard.edu

BIDMC Facility Director:

Joel Lawitts, PhD Instructor, BIDMC and Harvard Med. School
E-mail: jlawitts@bidmc.harvard.edu

The purpose of the Transgenic Core is:

• To provide a service facility that will utilize investigator-derived DNA constructs or investigator derived (or obtained) genetically modified embryonic stem cells to create founder transgenic and gene knockout or knockin mice (i.e mice bearing point mutants, lox-modified or ires/2A-Cre knockin alleles) that can be used to address questions relevant to obesity, diabetes and nutrition, including questions related to brain control of feeding, metabolism and reward.
• To provide consultation and training on all aspects of making and using genetically modified mice.

Core Services:

(with “charge-back” fees for BNORC members)

1. Generation of Transgenic Mice. DNA transgenic constructs, supplied by the investigator, are microinjected into fertilized one cell embryos. Tails from potentially transgenic founders are then supplied to the investigator for genetic analysis. Identified founders are then provided to the investigator. Transgenes can be injected into FVB/N or C57BL/6 mouse embryos. Other strains are available upon request. Charge Back Fee = $2,000 for FVB transgenics and $3,200 for C57BL/6 transgenics.
2. Gene Targeting – Generation of Targeted ES cells. DNA gene knockout constructs, supplied by the investigator, are electroporated into W4/129S6 embryonic stem cells (ES cell lines derived from other strains may be used following discussion with the Core). Drug resistant clones are selected, expanded and frozen down for subsequent use. Aliquots of cells from each clone are provided to the investigator for genetic identification of targeted clones. Charge Back Fee = $1,000.
3. Gene Targeting – Generation of Chimeric Mice from Targeted ES Cells. Targeted ES cells are injected into 3 day old embryos (blastocysts). Highly chimeric mice are then provided to the investigator for breeding. Targeted ES cell clones to be injected may have been generated by the BNORC Core (as above under #2), by the investigator’s lab, or by various high-throughput ES targeting consortiums (for example, EUCOMM - http://www.knockoutmouse.org/about/eucomm). Charge Back Fee = $650 for injection of 2 clones.
4. Cryopreservation of Mutant Mice. Fertilized one cell embryos are cryopreserved. Charge Back Fee = $3.50 per mouse embryo.
5. Generating Mice from Frozen Embryos. Embryos (frozen) from outside sources (after clearance from the BIDMC Animal Research Facility) or from the BNORC Transgenic Core Facility will be thawed and transferred into pseudopregnant recipients. Charge Back Fee = $550.
6. In Vitro Fertilization (IVF). B6D2F1 hybrid eggs will be inseminated with frozen or fresh sperm (pending import approval by the BIDMC Animal Research Facility). If fresh sperm is used, we will train investigators to cut out the tail of the epididymus. The resulting embryos will be transferred into pseudopregnant recipients. Charge Back Fee = $450.
7. Consultation. Consultation on all aspects of transgenic and gene knockout work is provided.

* Cost does not include per diems, shipping, etc.

Charges for Services

* Cost does not include per diems, shipping, etc.

After consulting with the Core Director, please use the link below to request Core services.

Click here to access the BNORC Core User Request Form