Genome Editing

Transgenic Mouse/Rat Production

To produce transgenic mice which will express a new gene function based on the DNA construct provided by the investigator. The transgene will be randomly incorporated within the germ line of the mouse and will be passed on to its progeny. The investigator will provide a DNA digest for purification by the facility. This DNA will be injected into the pronuclei of fertilized mouse eggs. Surviving embryos will be transferred to surrogate mothers and the tail tips from the pups born will be provided to the investigator for genotyping at weaning. A minimum of 3 positive founders will be delivered. 

CRISPR KO mice/rats

To produce CRISPR/cas9-mediated gene indels to generate knock-out mice or rats. The investigator will provide or purchase the CRISPR gRNA(s). TGEF will inject gRNAs and cas9 into pronuclei or cytoplasm of fertilized mouse eggs and transfer them to surrogate mothers. Tail tips from pups born will be collected at weaning and provided to the investigator for genotyping. Initial screening for indels can be difficult and consultation with or utilization of Purdue's Gene Editing Facility is encouraged. Positive founders will be transferred to the investigator's animal facility once identified.

CRISPR KI mice/rats

To produce CRISPR/cas9-mediated targeted gene inserts or edits in mice (including substitutions, cassette insertions and gene replacements). The investigator will provide or purchase insert DNA and CRISPR gRNA(s). TGEF will inject DNA, gRNAs and cas9 into pronuclei of fertilized mouse eggs and transfer them to surrogate mothers. Tail tips from pups born will be collected at weaning and provided to the investigator for genotyping. Initial screening for indels can be difficult and consultation with or utilization of Purdue's Gene Editing Facility is encouraged. Positive founders will be transferred to investigator's animal facility once identified.

Gene Targeting + Blast Injection

To produce targeted "knock­out" or "knock­in" mice. The DNA targeting construct will be provided by the investigator, designed to undergo homologous recombination with an endogenous mouse gene, deleting or replacing part of that gene such that it is functionally inactive or altered. The construct will be electroporated into embryonic stem cells and cultured in antibiotic selection media. Surviving, potentially targeted clones will be expanded and a duplicate set will be given to the investigator for screening. Clones harboring the correctly targeted gene will be expanded for injection into host mouse blastocysts. After injection, blastocysts are transferred to surrogate mothers and all chimeric pups born will be transferred at weaning to the investigator for breeding to confirm germ line transmission of the gene.