The purpose of the Genetically Engineered Mouse Facility (GEMF) is to provide technologically advanced and efficient mouse mutation resources to faculty members at the institution. To modify the genome, we perform direct DNA injection for standard transgenesis and CRISPR-mediated gene targeting. We also perform ES cell mutagenesis followed by blastocyst injection to generate modifications in the classical manner. Archiving and assisted reproduction, including embryo and sperm cryopreservation, in vitro fertilization and rederivation of mouse lines are all technologies supported by the GEMF.
The facility is fully equipped with the latest instrumentation and staffed with highly skilled personnel trained specifically in the production of mutant mice.
CRISPR/Cas9, ZFN and TALEN Injections
We are now performing CRISPR/Cas9, ZFN and TALEN injections. These unique methods of generating targeted mice hold great promise. Our initial CRISPR/Cas9 project was an exciting success, resulting in the publication "Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes. Yen ST et al., Developmental Biology 2014; 393(1):3-9." Since that publication, we have generated numerous CRISPR knock-out and knock-in mice. Please contact us for details on this procedure.
Sperm Cryopreservation and Assisted Reproduction
The GEMF uses the sperm cryopreservation and IVF protocols developed by Naomi Nakagata at the CARD Institute in Japan. With these protocols, we are routinely seeing 60-70% fertilization after thaw by C57BL/6 sperm, and 80-100% fertilization after thaw by FVB sperm.
The Dead Live Again
Did the last male of your critical line just die? If you put him in the refrigerator immediately (NOT the freezer), we can isolate sperm to recover the line for up to four days after death. In a previous procedure using a 6 to 8 month old male that had unfortunately expired, we ended up with a 79% fertilization rate. So don't give up hope. Call us at 713-745-1419 or 713-745-2654.