Director:Bradford Lowell, MD, PhD
Email: blowell@bidmc.harvard.edu

Facility Director:: Joel Lawitts, PhDPhD
Email: jlawitts@bidmc.harvard.edu

Asst. Core Director: Chen Wu, PhD
Email: cwu9@bidmc.harvard.edu

Asst. Core Director: Dong Kong, PhD
Email: Dong.Kong@tufts.edu


About the Core

The Transgenic Core generates transgenic and gene knockout mouse models relevant to obesity, diabetes and nutrition research. The Core also provides consultation on all aspects of making transgenic and gene knockout mice.

Activities:


-- 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 = $2,500 for FVB transgenics and $3,850 for C57BL/6 transgenics.

If repeat injections of a new DNA prep are required, because the prior injections failed to produce transgenic mice due to poor quality of investigator-provided DNA prep, this additional injection will cost $600* for FVB/N and $1,200* for C57Bl/6.

Guarantee: 4 founders. If  more than 50 pups were born without achieving 4 founders, injection can be repeated with above-mentioned DNA re-prep fee. 

-- CRISPR and “Easi-CRISPR” generation of gene knockout or gene knockin mice, mice bearing loxed alleles, Cre driver knockin mice, XFP knockin mice, etc. 

Reagents, including DNA constructs, guide RNAs and CAS9 protein supplied by the investigator, are microinjected into fertilized one cell embryos. The required amount of reagents is: 25ug CAS9, 15ug sgRNA, 3-5ug DNA (100 ng/ul minimum if in solution). Reagents may be presented to the Core in a lyophilized form (preferred), or reconstituted in culture grade water. Tails from potentially modified founders are then provided to the investigator for genetic analysis.  Identified modified mice are then provided to the investigator.  CRISPR reagents can be injected into FVB/N or C57BL/6 mouse embryos. Other strains are available upon request.

Charge Back Fee = $2,500 for FVB transgenics and $3,850 for C57BL/6 transgenics.  

Consultation and questions on all aspects of using CRISPR or Easi-CRISPR to make genetically modified mice is provide by Chen Wu (cwu9@bidmc.harvard.edu) and Brad Lowell (blowell@bidmc.harvard.edu). 

If repeat injections are required, no new reagents will be accepted. A re-injection fee of $600* for FVB/N and $1,200* for C57Bl/6 will be applied. Submission of new reagents will be billed as new project.

-- Gene Targeting – Generation of Chimeric Mice from Targeted ES Cells.  

Targeted ES clones 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 another Core, by the investigator’s lab, or by various high-throughput ES targeting consortiums (ie, EUCOMM – http://www.knockoutmouse.org/about/eucomm). 

Charge Back Fee for 1 day’s injection (either 1 or 2 ES clones) into black C57BL/6 embryos = $750.  

Charge Back Fee for 1 day’s injection (either 1 or 2 ES clones) into albino** C57BL/6 embryos = $900.

-- Cryopreservation of Mutant Mice.  

Fertilized one cell embryos are cryopreserved.

Charge Back Fee = $3.50 per mouse embryo

-- Generating Mice from Frozen Embryos.

Embryos (frozen) from outside sources (after clearance from the BIDMC Animal Research Facility) or from the BADERC Transgenic Core Facility will be thawed and transferred into pseudopregnant recipients.

Charge Back Fee = $550.

-- 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.

-- Consultation.  

Consultation on all aspects of transgenic and gene knockout work is provided.

 

* Cost does not include per diems, shipping, etc. 
** Albino C57BL/6 embryos facilitate visual detection of chimerism and germline transmission when black C57BL/6 ES clones are being injected. Albino embryos are derived from C57BL/6J-Tyrc-2J mice.


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