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. Modifications to the genome utilizing direct DNA injection and ES cell mutagenesis, cryopreservation, in vitro fertilization and rederivation of mouse lines are all technologies supported by the facility.
The facility is fully equipped with the latest instrumentation and staffed with highly skilled personnel trained specifically for the production of mutant mice.
New Sperm Cryopreservation and IVF Protocols
The GEMF is now using new 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.
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 first CRISPR/Cas9 project was an exciting success, with at least one mouse exhibiting a double-allele knock out at the single cell stage, and other mice with double knock outs at later stages to generate mosaics. The co-injection of mRNA with plasmid DNA is a technique that the GEMF has already used successfully to generate transposon-mediated transgenic mice.
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. We did this recently with a 6 to 8 month old male and ended up with a 79% fertilization rate. So don't give up hope. Call us at 713-745-1419 or 713-745-2654.