Rippel Mouse ES Cells and Transgenics Facility

Room E18-218
Ph. 617-452-3658
FAX 617-324-3912

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This shared resource of the Koch Institute provides fee-for-service support to all MIT investigators who utilize novel mouse models to study human diseases such as cancer.  
     
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Due to parameters established by our funding, we are NOT able to provide any services to non-MIT investigators. If you are interested, the following Transgenic Facilities seem to provide ES cell services for other academic institutions

For an explanation of how fees are set, see the bottom of this page.

  1. Assistance with transgenic vector design

    Fee: None

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

  2. Distribution of plasmids useful as transgenic vector backbones

    Fee: None

    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
      (Addgene 13445 for map and sequence)
  3. Gene Targeting in ES cells

    Fee per targeting with more than 96 colonies picked:
    $1500 for KI, $3000 for non-KI
    Fee per small targeting, picking 96 colonies or fewer:
    $1000 for KI, $2000 for non-KI

    To make a knockout/knockin mouse with the Rippel Transgenic Facility, 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.

  4. Culture of ES cells and preparation for blastocyst injection

    Fee : $150 for KI, $225 for non-KI

    Targeted ES cells or ES cells containing gene traps will be expanded, tested for pathogens (if needed, added cost of $450), and prepared for injection into appropriate blastocysts.

  5. 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. The test itself costs $450.
    Fee: $50 for KI, $75 for non-KI

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

    Fee for injection of DNA or virus into fertilized eggs:
    $1200 for KI, $1650 for non-KI
    Fee for injection of ES cells into B6 or Balb/c (*) diploid blastocysts:
    $1200 for KI, $1650 for non-KI
    Fee for injection of F1 hybrid ES cells into tetraploid blastocysts:
    $1800 for KI, $2400 for non-KI
    Fee for each additional recipient mouse: $35 for KI, $45 for non-KI

    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. 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.

    * A surcharge will be added to the cost for investigators who do not use the two standard strains of mice (either C57BL6 or Balb/c) for donors. The surcharge will cover the purchase cost of the mice as well as per diem and colony management. Please contact Kirk Maurer at DCM for special requests and for an estimate of the costs.

  7. Provide tissue-specific Cre mouse lines

    Fee per mouse: $30 to $300, depending on strain and affiliation

    A collection of useful strains of mice are maintained by the Rippel Transgenic Facility and are available to MIT researchers. These strains include approximately 30 Cre mice, FLPe 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.

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

    Hands-on training, mouse breeding assistance, etc is
    $55 per hour for KI and $82.50/hr for non-KI

    Training options include, but are not limited to:

    • Generation of MEFs
    • Isolation of mouse embryos for histology or other analysis
    • Isolation and culture of blastocysts
    • Generation of ES cell lines
    • Culture of ES cells
    • Differentiation of ES cells to specific tissue type
    • Karyotyping
    • Optimizing breeding conditions for mice
  9. Provide reagents needed for ES cell or other issue culture

    Vials of 3x10`6 irradiated MEFs for use as feeders -$6 for KI, $9 for non-KI
    Vial of p1 MEFs - $50/vial for KI, $75 for non-KI
    Vial of germline-proven ES cells - $250/vial for KI, $375 for non-KI
    Media, Serum, and ESGRO (at cost) for ES cell culture

Other services are also available on a limited basis, please inquire to Aurora if you are interested.

Explanation of how fees are set

Some support for salaries of the Transgenic Facility Staff is provided by the National Cancer Institute Cancer Center Support Grant to the MIT Center for Cancer Research. Therefore, services to KI members have a subsidized rate.

For Gene Targeting, the base $1500 covers all media, cells, plastics and other supplies needed for the procedure as well as a small amount of salary. Each targeting requires 25-30 hours of labor, which is reflected in the increased price for non-KI members. Targetings with fewer colonies picked require fewer materials and less labor.

Other fees for cell culture have been similarly established to cover materials and labor.

Most of the Cre lines in the repository are available from Jackson Labs, at a cost ranging from $84-$475 per mouse, with an average cost of $174/mouse plus cost of processing and shipping. We have priced our mice to be less expensive than JAX mice and require no shipping fees. We have also backcrossed some of the Cre lines to a more useful background (lines for immunological studies onto C57B6, for example). Our fee offsets the costs of cage perdiems, colony maintenance and genotyping.

 
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updated 5/18/08
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