For pricing and an explanation of how fees are set, see pricing page.
To request a service, please see Core Access page.
Please begin with the "Beginner's Guide to Gene Targeting," written by Aurora Burds Connor under the Methods section, but Aurora (firstname.lastname@example.org) is also happy to help with individual questions for your construct.
We currently have vectors containing combinations of:
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.
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.
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 "Core Access."
To Request a Pronucler injection (PNI) of DNA, CRISPR/Cas9 components or virus into fertilized mouse eggs, please click here.
For CRISPR PNI, we have stock of Cas9mRNA from TriLink BioTechnologies available. Please email us if you are intrested in an aliquot.
In addition to the injections services offered through the support of the KI, other services such as cryopreservation of sperm and embryos, IVF, and mouse genotyping are also available. Please see the DCM website (MIT Certificates Required) for more details and for the DCM forms to request these services.
A collection of useful strains of mice are maintained by this Facility and are available to MIT researchers. These strains include Cre mice, FLP mice, Cas9 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.
Training options include, but are not limited to:
Other services are also available on a limited basis, please inquire to the ES Core Staff (email@example.com) if you are interested.