Penn Gene Targeting

Overview: Penn Gene Targeting (PGT) provides a full DNA - to - mouse service available to all Penn researchers and all academic researchers outside. This includes design of BAC-derived targeting vectors by PGT in collaboration with the Client Lab and construction of the vectors by PGT in collaboration with the DNA Sequencing Facility. Finished targeting vectors are electroporated into one of several 129 or C57BL6 mouse ES cell lines and selected with suitable drugs, depending on the vector. The client lab or PGT then screens ES lines by Southern/PCR and together with PGT identifies the correct recombinant clones. Recombinant clones are tested by PGT for cytogenetic integrity and expanded for injection into blastocysts. PGT coordinates timing of the expansion with the Transgenic and Chimeric Mouse Facility, which performs the injection into mouse blastocysts and produces the chimera.

Prices: Prices are about one third to one half below those of commercial services for researchers of the University of Pennsylvania. In fact, even for outside academic researchers prices are still about one third below average commercial services.

Funding: PGT has been supported by the NIH-sponsored Knockout Mouse Project (KOMP) and the Penn Diabetes and Endocrinology Research Center (DERC).

Rates of germline transmission for our ES cell lines. PGT achieves high rates of germline transmission for its ES cell lines. In particular, PGT has stringently tested improved growth conditions for C57BL6 ES cells and more than 30 independently targeted C57BL6 ES clones were generated and tested for germline transmission. As a result PGT routinely achieves rates of germline transmission for this desirable but technically more challenging mouse background that are comparable to the rates for 129 ES cells. For detailed and extensive documentation on germline transmission rates of our C57BL6 ES cell lines please contact Dr. Tobias Raabe.

Outstanding Results: Since its formal establishment in 2005, PGT and the collaborating DNA Sequencing and Chimeric Mouse cores have successfullytargeted more than 40 genes for clients, resulting so far in more than 32 final KO mice with the rest of the projects in various stages of completion. Several of these mice have now being published in journals and/or abstracts (please contact Dr. Raabe for details).