Production of transgenic mice by DNA injection into pronuclei of mouse oocytes
This service includes hormone preparation of female donors, mating of females with stud males, harvesting of fertilized ova, microinjection of foreign DNA, preparation of surrogate females by mating with vasectomized males, and oviduct transfer of injected eggs into pseudopregnant foster females. Five to eight transfers of approximately 20 injected eggs each will be performed.
The Transgenic and Reproductive Technology Center personnel weans 3 weeks old pups and submits foster mothers for blood serology. After results arrive negative for all tested pathogens, litters will be transferred into the investigator’s room. Tail tipping can be performed at the investigator’s request for an additional charge of $40 by Transgenic and Reproductive Technology Center personnel or ordered by the investigator through LARC services. The investigator is responsible for timely reports of genotyping results to the Transgenic and Reproductive Technology Center. If results are not reported on time (2 weeks), all cages with litter will be transferred into the investigator’s room. If the number of founders is less than three, DNA (freshly prepared) will be injected for free once.
Procedure Outline
- The investigator submits DNA for purification to the CRISPR and Genome Editing Center. DNA should be prepared for purification according to GTF protocol.
- Gene Targeting Facility personnel purifies the transgene.
- Researcher registers the request to inject the DNA construct on the GTF website.
In order to obtain a password for the database, please contact Flora at x8650. Microinjection of the DNA construct is done on a “first-come, first-served” basis.
DNA microinjection into pronucleus of the mouse oocyte