Services

For pricing and an explanation of how fees are set, see pricing page.

To request a service, please see Forms page.

Assistance with transgenic vector design

Please begin with the "Beginner's Guide to Gene Targeting," written by Aurora Burds Connor under the Methods section, but Aurora (escore@mit.edu) is also happy to help with individual questions for your construct.

Distribution of plasmids useful as transgenic vector backbones

We currently have vectors containing combinations of:

  • Neo cassettes
  • TK cassettes
  • DTA cassettes
  • pPGKneoF2L2dta- Neo flanked by both Frt and LoxP sites, with Diptheria Toxin A
    (Addgene 13445 for map and sequence)

Gene Targeting in ES cells

To make a knockout/knockin mouse with the Mouse ES Cell and Transgenic Facility, MIT investigators provide the targeting construct as well as proof of a viable screening strategy, and the facility will electroporate the DNA into ES cells, perform drug selection, subclone ES cell colonies, and prepare genomic DNA for screening. The ES cell facility then coordinates with the Division of Comparative Medicine for the injection of targeted clones into blastocysts, which occurs in a pathogen-free barrier animal facility. (click here for a flowchart and timeline)

Standard targetings produce only 1%-2% positive clones, so it is necessary to pick approximately 300 colonies to increase the likelihood of obtaining enough correctly targeted cells. Some targetings are directed to loci with high recombination rates (ROSA26 at 50%-75%, for example) and these require fewer colonies for success.

Culture of ES cells and preparation for blastocyst injection

Targeted ES cells or ES cells containing gene traps will be expanded, tested for pathogens, and prepared for injection into appropriate blastocysts.

Preparation of cells for pathogen testing (pathogen tests are required for all imported cell lines before they will be injected). Up to 5 lines can be pooled together for testing.

Injection of DNA, ES cells, or virus into pre-implantation embryos

MIT's Division of Comparative Medicine (DCM) and the KI pooled their efforts, resources, and personnel in the late 1990's to provide a full range of services for the needs of the MIT mouse community. 

Requests for ES Cell-based services are processed through iLab.  Instructions and information about iLab are listed under "Forms."

Requests for Pronucler injection of DNA (PNI) or injection of virus are processed by DCM.  In addition to the injections services offered through the support of the KI, other services such as cryopreservation of sperm and embryos, IVF, and genotyping are also available. Please see the DCM website (MIT Certificates Required) for more details and for the DCM forms to request these services.

Provide tissue-specific Cre mouse lines

A collection of useful strains of mice are maintained by this Facility and are available to MIT researchers. These strains include a dozen Cre mice, FLP mice, reporter mice, and important wild type strains that are not otherwise available for direct import to the 68N facility. Please click for a list and description of mice currently available and for detailed fees and ordering instructions. As with all our services, we are not able to distribute the mice outside of MIT.

Provide methods (link on left), assistance and training in embryo and mouse analysis

Training options include, but are not limited to:

  • Culture of ES cells or iPS cells
  • Generation of new ES cell lines
  • Isolation and culture of blastocysts
  • Generation of MEFs
  • Isolation of mouse embryos for histology or other analysis
  • Differentiation of ES cells to specific tissue type
  • Optimizing breeding conditions for mice

Provide reagents needed for ES cell or other issue culture

  • Vials of 3x10`6 irradiated MEFs for use as feeders
  • Vial of p1 MEFs
  • Vial of germline-proven ES cells
  • Media, Serum, and ESGRO for ES cell culture

 

Other services are also available on a limited basis, please inquire to the ES Core Staff (escore@mit.edu) if you are interested.