Facilities
Transgenic and genome editing
The Transgenic and Genome Editing Facility creates and supports transgenic and gene-edited mouse and rat models to advance research on development, physiology, metabolism, and human diseases, including cancer.
About
In addition to a fully equipped microinjection and embryo laboratory, the TGEF contains a dedicated tissue culture laboratory and strictly maintained barrier containment facility housing both mice and rats. The facility is fully equipped for microinjections, microsurgery, zygote electroporation, gene targeting and tissue culture, and together with Purdue’s Gene Editing Facility and MilliporeSigma’s CRISPR Core Partnership can provide expert consultation on the design and execution of experiments.
Our services include the creation of transgenic, knock-out, knock-in and gene-edited mice and rats. We offer pronuclear injection of DNA into zygotes, gene targeting in ES cells and pronuclear or cytoplasmic injection as well as zygote electroporation of CRISPR/cas9 for the creation of single or multiple gene modifications, deletions, insertions and replacements.
We also offer many assisted reproductive technologies including cryopreservation of embryos and sperm; rederivation for the importation or “clean-up” of infected mice or rats; in vitro fertilization (IVF) to overcome poor reproductive performance due to age or other environmental or physiological factors, and a speed expansion service using IVF to rapidly generate large numbers of genetically engineered mice.
For members on the Purdue campus, we also offer a limited number of C57BL/6N mice for sale from our in-house breeding colony.
Capabilities
Transgenic mouse/rat Production
Our facility helps researchers produce transgenic mice that will express a new gene function based on the DNA construct provided by the investigator. The transgene is randomly incorporated within the germ line of the mouse and passed on to its progeny. The investigator provides a DNA digest for purification by the facility. This DNA is injected into the pronuclei of fertilized mouse eggs. Surviving embryos are transferred to surrogate mothers and the tail tips from the pups born are provided to the investigator for genotyping at weaning. A minimum of three positive founders are delivered.
CRISPR KO mice/rats
The TGEF produces CRISPR/cas9-mediated gene indels to generate knock-out mice or rats. The investigator provides or purchases the CRISPR gRNA(s). TGEF injects gRNAs and cas9 into pronuclei or cytoplasm of fertilized mouse eggs and transfers them to surrogate mothers. Tail tips from pups born are collected at weaning and provided to the investigator for genotyping. Initial screening for indels can be difficult; consultation with or utilization of Purdue’s Genomics and Gene Editing/Cell Line Development Facility is encouraged. Positive founders are transferred to the investigator’s animal facility once identified.
CRISPR KI mice/rats
Our facility produces CRISPR/cas9-mediated targeted gene inserts or edits in mice (including substitutions, cassette insertions and gene replacements). The investigator provides or purchases insert DNA and CRISPR gRNA(s). TGEF injects DNA, gRNAs, and cas9 into pronuclei of fertilized mouse eggs and transfers them to surrogate mothers. Tail tips from pups born are collected at weaning and provided to the investigator for genotyping. Initial screening for indels can be difficult; consultation with or utilization of Purdue’s Genomics and Gene Editing/Cell Line Development Facility is encouraged. Positive founders are transferred to the investigator’s animal facility once identified.
Gene targeting + blast injection
Our facility produces CRISPR/cas9-mediated targeted gene inserts or edits in mice (including substitutions, cassette insertions and gene replacements). The investigator provides or purchases insert DNA and CRISPR gRNA(s). TGEF injects DNA, gRNAs, and cas9 into pronuclei of fertilized mouse eggs and transfers them to surrogate mothers. Tail tips from pups born are collected at weaning and provided to the investigator for genotyping. Initial screening for indels can be difficult; consultation with or utilization of Purdue’s Genomics and Gene Editing/Cell Line Development Facility is encouraged. Positive founders are transferred to the investigator’s animal facility once identified.
Assisted reproduction
Rederivation by embryo transfer
The TGEF allows mice or rats from conventional housing or those with known or suspected pathogens to be brought into a barrier facility by embryo transfer. “Dirty” male mice/rats are held in quarantine where they are mated with donor females. Embryos from these matings are washed and transferred to clean surrogate mothers in our barrier facility. At weaning, surrogate mothers are bled and serum prepared and shipped for serological testing. Once the pups are confirmed pathogen free, they are transferred to the investigator’s animal facility.
Embryo cryopreservation
Our facility can preserve valuable lines of mice and rats for security and to avoid costly long-term maintenance of breeding colonies. Valuable germplasm is cryopreserved as embryos and stored in straws immersed in liquid nitrogen. The investigator provides 7-8 males who are mated multiple times with wild type females supplied from TGEF’s in-house colony. Investigator-provided females may be utilized for an additional fee and after consultation with the TGEF director on breeding strategy. 300 embryos (200 for rats) will be collected from the females and frozen. Frozen embryos can later be thawed and transferred into surrogate mothers when they are needed, either for research or to replace contaminated or infected lines. There is a yearly storage fee for each line stored in TGEF’s duplicate cryobanks.
Sperm cryopreservation
<text> The TGEF can preserve sperm from valuable lines of mice for security and to avoid costly long-term maintenance of breeding colonies. Valuable germplasm is collected from two males, cryopreserved and stored in straws immersed in liquid nitrogen. Sperm can later be thawed and used for IVF to repopulate the colony when needed, either for research or to replace contaminated or infected lines. There is a yearly storage fee for each line stored in TGEF’s duplicate cryobanks. At this time, sperm cryopreservation is available only for mice.
In vitro fertilization
Our facility uses IVF to help investigators overcome breeding problems in valuable mice. Transgenic or otherwise valuable mice frequently encounter breeding problems related to genetic manipulation, in-breeding or simply age. This technique allows investigators to save lines of mice which would otherwise be lost. Sperm is collected from the mouse in question and incubated with unfertilized oocytes from donor females. Successfully fertilized eggs are then transferred to surrogate mothers who give birth to offspring of the original infertile male. This technique is also used to regenerate lines from frozen sperm. At this time, only available for mice.
Speed (embryo) cryopreservation
Same as embryo cryopreservation as described above, but utilizing IVF and only two males, to quickly generate the 300 embryos necessary to secure the line. At this time, speed (embryo) cryopreservation is available only for mice.
Speed expansion
The TGEF can rapidly produce a large number of mice using IVF. Sperm is collected from two or three male mice of the line required and incubated with oocytes from wild type donor females. Successfully fertilized eggs are transferred to surrogate mothers who give birth to offspring of the original male. At this time, speed expansion is available only for mice.
Rates – Mouse
| Service | PCCR Member Rate | Internal Rate | External Rate |
|---|---|---|---|
| Genome Editing | |||
| Transgenic Mouse Production | $2,575 | $6,230 | $9,781 |
| Additional TG Injection Session | $715 | $1,375 | $2,159 |
| CRISPR KI mice | $2,760 | $7,700 | $12,089 |
| CRISPR KO mice | $2,095 | $5,170 | $8,117 |
| CRISPR Validation | $60 | $260 | $408 |
| Additional CRISPR Injection Session | $750 | $1,725 | $2,708 |
| Gene Targeting + Blast Injection | $7,500 | $18,950 | $29,752 |
| Blast Injection of ES Cells | $4,900 | $11,660 | $18,306 |
| Assisted Reproduction | |||
| Rederivation by Embryo Transfer | $235 | $410 | $644 |
| Embryo Cryopreservation | $470 | $1,375 | $2,159 |
| Speed (Embryo) Cryopreservation | $885 | $1,740 | $2,732 |
| Sperm Cryopreservation | $130 | $270 | $424 |
| In Vitro Fertilization | $335 | $900 | $1,413 |
| Speed Expansion | $750 | $1,840 | $2,889 |
| Cryo Bank per Line charge | $18 | $22 | $34 |
| Tissue Culture | |||
| Chromosome Counting | $100 | $360 | $565 |
| Mouse Embryonic Fibroblast Production | $450 | $970 | $1,523 |
| Husbandry | |||
| Production Colony | $14 | $14 | N/A |
| Mouse Colony Set Up | $100 | $130 | N/A |
| Animal Shipment (Domestic) + Packaging at cost | $140 | $140 | $220 |
| Animal Shipment (International) + Packaging at cost | $235 | $235 | $369 |
| Miscellaneous | |||
| Animal Labor Rate | $47 | $47 | $74 |
| Lab Labor Rate | $38 | $38 | $60 |
| Consultation Rate | $80 | $80 | $126 |
Rates – Rat
| Service | PCCR Member Rate | Internal Rate | External Rate |
|---|---|---|---|
| Genome Editing | |||
| Rat Transgenic Production | $3,810 | $8,410 | $13,204 |
| Additional TG Injection Session | $900 | $2,010 | $3,156 |
| CRISPR KI Rats | $4,000 | $9,580 | $15,041 |
| CRISPR KO Rats | $2,600 | $6,040 | $9,483 |
| CRISPR Validation | $75 | $280 | $440 |
| Additional CRISPR Injection Session | $1,000 | $2,170 | $3,407 |
| Assisted Reproduction | |||
| Rederivation by Embryo Transfer | $420 | $610 | $958 |
| Embryo Cryopreservation | $990 | $1,900 | $2,983 |
| Cryo Bank per Line charge | $18 | $22 | $34 |
| Husbandry | |||
| Animal Shipment (Domestic) + Packaging at cost | $140 | $140 | $220 |
| Animal Shipment (International) + Packaging at cost | $235 | $235 | $369 |
| Miscellaneous | |||
| Animal Labor Rate | $47 | $47 | $74 |
| Lab Labor Rate | $38 | $38 | $60 |
| Consultation Rate | $80 | $80 | $126 |