Shared Facilities

Transgenic & Knock-Out Mouse

Policies & Procedures

Transgenic & Knock-Out Mouse Core service charges are as follows. See below for transgenic and knock-out mouse production procedures.

Transgenic & Knock-Out Mouse Core Service Charges
(Effective July 2015)
Service Pricing
Pronuclear injection into C3HeB/FeJ or C57/Bl6J embryos $5,835
ES cell injection into C57Bl/6J blastocysts $4,197
Sperm cryopreservation (with verification) $1,791
Sperm cryopreservation (without verification) $662
Cryopreservation of IVF embryos $994
In vitro fertilization with DBA/2J, C3HeB/FeJ or C57Bl/6J ova and embryo transfer $1,697
Embryo thaw and transfer $897
Cryo storage $70/line/yr.
Procedures

1. For pronuclear injection, investigators provide the DNA construct to the core. The core then: 

  • Gel purifies traditional transgene constructs
  • Harvests single cell embryos
  • Microinjects the embryos with the transgene or CRIPSR construct
  • Transplants the microinjected embryos into pseudo-pregnant females
  • Performs Cesarean sections as required
  • Provides the resulting pups at weaning age to the investigator for analyses

The core guarantees 10 births per round pronuclear injection when using C3HeB/FeJ embryos (historical average is 15 births per round).
The core guarantees 5 births per round pronucear injection when using C57Bl/6J embryos (historic average is 14 births per round).

2. For single ES cell line blastocyst injection, investigators provide the ES cell line to the core.  The core then:  

  • Amplifies the ES cell ine
  • Harvests C57Bl/6J blastocysts
  • Injects the ES cells into the blastocyst
  • Transplants the injected blastocysts into pseudo-pregnant females
  • Performs Cesarean sections as required
  • Culls the resulting chimeric animals

The core guarantees 6 births per round of ES cell line blastocyst injection.

3. For sperm cryopreservation with verification, investigators provide a minimum of two male mice.  The core then:

  • Harvests sperm from the donor
  • Produces 50 straws of frozen sperm
  • Verify successful freezing via in vitro fertilization using the thawed sperm and donor eggs from the relevant strain of mice; verification entails development of  2-cell embryos following in vitro fertilization

The verification is the guarantee for this service.

4. For sperm cryopreservation (without verification), investigators provide a minimum of two male mice.  The core then:

  • Harvests sperm from the donor
  • Produces 40 straws of frozen sperm

The core guarantees mobile sperm upon test thawing. 

5. For cryopreservation of IVF embryos, investigators provide either frozen sperm samples or a minimum of two living make mice.  The core then:

  • Thaws the frozen sperm or harvests fresh sperm from the donor
  • Performs in vitro fertilization
  • Cultures embryos to 2-cell stage
  • Cryopreserves the embryos

The core guarantees generation of 2-cell frozen embryos (yield is strain dependent: C3HeB/FeJ, 30 embryos;  DBA/2J, 40 embryos; C57Bl/6J, 15 embryos); viability confirmed via culture of a thawed sample to 4-cell stage.

6. For in vitro fertilization with DBA/2J, C3HeB/FeJ or C57Bl/6J ova and embryo transfer, investigators provide either frozen sperm samples or a minimum of two living make mice.  The core then:

  • Thaws the frozen sperm or harvests fresh sperm from the donor
  • Performs in vitro fertilization
  • Cultures embryos to 2-cell stage
  • Transplants the 2-cell embryos into pseudo-pregnant females
  • Performs Cesarean sections as required
  • Provides the resulting pups at weaning age to the investigator for analyses

In the event that that no births are obtained, a second round (optimized based on the results from the first round) will be performed for no additional charge.   Subsequent efforts will require additional billing.

7. For embryo thaw and transfer, investigators provide the frozen embryos.  The core then:

  • Transplants the 2-cell embryos into pseudo-pregnant females
  • Performs Cesarean sections as required
  • Provides the resulting pups at weaning age to the investigator for analyses

In the event that that no births are obtained, a second round (optimized based on the results from the first round) will be performed for no additional charge.   Subsequent efforts will require additional billing.

Notes:

  • All charges and guarantees are based on the costs and yields encountered over the past 5 years of Core activity.
  • Cryopreservation activities are based on a good faith agrement between the investigators and the Core.  Thus far the Core has not encountered any difficulties in the recovery of cryopreserved samples which it has generated.  However we cannot guarantee that all samples can be recovered following long-term storage.