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Transgenic Mouse Facility

Transgenic Mouse Facility

The Transgenic Mouse Facility in the Division of Animal Resources offers mutant mouse model production services to investigators for their research projects. Services in the facility include transgenic mouse production by pronuclear microinjection and knockout and knockin mouse production using both CRISPR gene editing and traditional mouse ES cell gene targeting, mouse line cyropreservation, and rederivation.

Services

DNA Microinjection

CRISPR-mediated gene edited mice or transgenic founder mice are generated by injecting CRISPR reagents or a transgene into mouse zygotes. Injected mouse embryos will be implanted into foster females to carry them to term. The gestation period of a mouse is approximately 20 days. Pups will be transferred to an investigator's protocol for genotyping at 2 to 3 weeks of age. 

To request this service, investigators should: 

  • Have their project approval from both Biological Safety Committee (IBC) and IACUC.
  • Arrange an appointment with Director Fuming Pan (244-0649 or fpan@illinois.edu) to initiate DNA microinjection.
  • Submit a service request form.

Note: For transgenic mouse production, provide the facility with 20ug of digested plasmid DNA at 0.5ug/ul or higher with a picture of the DNA prepared for fragment isolation.

Gene Targeting

A targeting vector will be introduced into ES cells via electroporation. ES cells will then be grown in appropriate selection media. Around 200 ES cell clones will be picked up, duplicated, and given to the investigator for analysis. Positive ES clones will then be expanded, frozen, and kept in the facility for subsequent blastocyst injection to generate chimeric mice. 

To request this service, investigators should: 

  • Schedule an appointment with Director Fuming Pan (244-0649, fpan@illinois.edu) to discuss the targeting strategy.
  • Have the DNA construct approval from the IBC.
  • Submit a Service Request Form.

Note: Investigators should provide at least 200ug linearized DNA at 0.5ug/ul or higher to the facility.

ES Cell Microinjection

Chimeric mice can be generated by microinjection of targeted ES cells into 3.5 days old mouse embryos. The facility will perform the microinjection and transfer injected blastocysts into foster families. Chimeric mice will be transferred to an investigator’s protocol after weaned. Since mouse ES cells are pluripotent and can contribute to all cells in an animal, including gametes, genetic alterations introduced into the ES cells can be propagated in the next generation to form a stable mutant mouse line. 

To request this service, investigators should: 

  • Have their project approval from both IBC and IACUC.
  • Arrange an appointment with Director Fuming Pan (244-0649, fpan@illinois.edu) to initiate ES cell microinjection.
  • Submit a service request form.

Note: If the investigator already has ES cell clones, please provide the facility with ES cells and specified protocols. The facility will provide at least two chimeric mice per knockout construct.

Embryo Cyropreservation

Mouse embryos can be cryopreserved to prevent the accidental loss of a valuable mouse line or to discontinue the maintenance of a mouse stock. The Transgenic Mouse Facility will isolate and cryopreserve 8-cell embryos from superovulated female mice using the slow cooling method. The investigator will be responsible for providing a sufficient number of mice. Usually, about 200 embryos per mouse line will be cryopreserved. 

To request this service, investigators should: 

  • Arrange an appointment with Director Fuming Pan (244-0649, fpan@illinois.edu) to discuss the project.
  • Complete and submit the service request form.

Note: The investigator will need to have space to store the cryovials once the project is finished.

Rederivation

The Transgenic Mouse Facility will perform experiments to clean a mouse line from pathogen infections. The procedure will include superovulating, mating, and collecting fertilized one-cell embryos. The investigator is responsible the mouse purchase. Females are super-ovulated and bred to males of the line to be rederived. Embryos are collected and pooled, washed to further reduce the risk of pathogen transfer, and then surgically transferred, using aseptic technique, into the reproductive tract of pseudopregnant SPF females. Upon weaning of the pups, DAR will submit surrogate dams for diagnostic testing, including screens for bacterial and viral pathogens. 

To request service from Transgenic Mouse Facility, investigators should: 

  • Schedule an appointment with Director Fuming Pan (244-0649, fpan@illinois.edu) to discuss the rederivation project.
  • Provide 3 to 4 proven breeder males and 3 to 4 females, both aged 3 to 4 weeks, for each line to be rederived.
  • Submit a Service Request Form.

Note: Investigators are responsible for the per diem from a quarantine room and any related tests.

Embryo Recovery

The Transgenic Mouse Facility can retrieve mouse lines from both fresh and frozen embryos. The procedures include thaw and collection of mouse embryos and surgically transferring them into foster females using aseptic techniques. 

To request these services, investigators should: 

  • Schedule an appointment with Director Fuming Pan (244-0649, fpan@illinois.edu) to discuss the embryo recovery project.
  • Provide at least 40 embryos with health documents for each line to be recovered.
  • Submit a Service Request Form.

Note: The Investigator is responsible for per diem from the date of the embryo transfer.

Miscellaneous Services

The Transgenic Mouse Facility staff can train or assist individual mouse users in aspects of mouse colony breeding, such as weaning pups, extracting tail DNA, setting up timed mating, dissecting mouse embryos, and handling mouse ES cell cultures. These services are contingent upon the availability of time, equipment, animal health, and other factors. 

To request these services, investigators should: 

  • Schedule an appointment with Director Fuming Pan (244-0649, fpan@illinois.edu) to discuss this project.
  • Submit a Service Request Form.

Pricing

Service Campus Price Off-Campus Price
DNA Microinjection $5,210 $5,731
Gene Targeting $6,560 $7,216
ES Cell Microinjection $5,340 $5,874
Embryo Cryopreservation $1,790  
Rederivation $1,820  
Embryo Recovery $2,040  
Miscellaneous Services $40/hr  

Note: Additional charges include embryo donor and recipient purchase for each project

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