JAX® Microinjection Services

The Jackson Laboratory offers both DNA microinjection and ES cell microinjection services to facilitate the creation of genetically modified mice. The development of transgenic mouse technology has roots at The Jackson Laboratory1 and we have successfully created thousands of genetically modified mice.

DNA Microinjection Service

Our Transgenic Services Program will prepare your DNA for microinjection according to well-established and proven protocols and microinject it into 250 embryos derived from the C57BL/6J strain (Stock Number 000664).  In most cases, greater than 50 mice will be produced for subsequent testing for DNA integration.  In our experience, approximately 95% of all DNA microinjection projects yield transgenic founder mice. (However, due to the inherent variables associated with the transgenic animal production process, we cannot guarantee the production of transgenic mice from the DNA you provide.) All resulting mice will be shipped to you after weaning and will be specific pathogen-free (a health report will accompany the shipment of your animals).

Customers need to provide:

  1. At least 50ug of purified plasmid DNA of sufficient purity for restriction digest, or 50ug of purified BAC DNA.
  2. A map of the plasmid with required restriction enzyme sites denoted.
  3. For plasmids that require linearization and/or gel purification, an image of the digested plasmid run on an agarose gel with a defined DNA ladder.
  4. Information regarding expected phenotype (if any), or any other information that may influence the outcome of the service.
Note: We cannot accept transgenic service projects that will require laboratory or animal room procedures at the BSL2/ABSL2 level or higher.  For example, we will not make transgenic mice from lentiviral or adenoviral vectors, either by infection or DNA microinjection.

ES Cell Microinjection Service

The creation of mutant mice by gene-targeting in embryonic stem (ES) cells is most commonly used to ablate (knock out) gene function. The introduction of genetically modified ES cells into mouse embryos is a well-established technique that has been used to create thousands of mutant mouse lines.  It can also be used to introduce a specific mutation into the mouse genome.

Our ES cell microinjection program will microinject strain 129 or C57BL/6J ES cells (gene targeted in your laboratory) into 100 C57BL/6J (Stock Number 000664) or B6(Cg)-Tyrc-2J/J (Stock Number 000058) embryos, respectively. Any resulting mice will be scored for the degree of coat-color chimerism. All chimeric mice born will be shipped to you and will be specific pathogen-free.  Note that we cannot guarantee that your ES cells will contribute to embryonic development and result in chimeric offspring, or that any chimeric mice created will ultimately produce offspring of the desired genotype. In our experience approximately 90% of projects yield coat color chimeric mice and greater than 20 mice are produced.  

We will test a sample of your ES cells for the presence of common mouse pathogens prior to injection.  Only ES cell samples that are proven free of mouse pathogens will be microinjected into host embryos for the production of chimeric mice.

Customers need to provide:

  1. At least two frozen vials of ES cells containing no less than 6 million cells each.
  2. Appropriate background information on the cells, such as the parental cell line, special culture conditions if applicable, expected phenotype if any, and any other information that may influence the outcome of the service.
Note that a key component to successful mutant mouse production is the ability of the ES cell line to colonize the developing host embryo and contribute to the germline of the resulting mice.  The ES cells used in this technique must be maintained properly and have a normal karyotype.  Mouse ES cells that do not have a normal karyotype (complete and correct complement of chromosomes) will not produce germline competent mice, or at best, produce germline competent mice at a very low frequency. 

References

  1. Wagner TE, Hoppe PC, Jollick JD, Scholl DR, Hodinka RL, Gault JB. 1981. Microinjection of a rabbit beta-globin gene into zygotes and its subsequent expression in adult mice and their offspring. Proc Natl Acad Sci USA. 78:6376-6380.

Advantages

  • Leverages our extensive model creation experience
  • Both DNA and ES cell microinjection are available
  • All resulting mice are shipped Specific Pathogen Free (SPF)

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