Core Description: The Penn Gene Targeting Service (PGT) is run by the Penn Gene Targeting Core and Laboratory in collaboration with two other Penn Cores (see below) and has the capability to generate from scratch almost any type of genetically modified mouse. This includes complete knockout of any gene, tissue-specific and/or drug-inducible knockout of any gene, targeted mutagenesis of any region of any gene (knockin), tissue-specific activation of an endogenous gene using loxP flanked stop cassettes, and tissue-specific and/or drug-inducible transgene expression from defined loci. For details regarding services and prices please consult the PGT home page of the Department of Genetics and/or directly contact Dr. Tobias Raabe.
The Client lab does not need to be involved in any experimental step and only needs to breed the resulting mice. However, if desired, participation of the client lab in almost any experimental step is possible.
PGT has an outstanding record of germ line transmission and so far over 32 targeted mice - more than 12 in the desirable C57BL6 background - have been made, starting either with molecular design and construction of the targeting vector or with electroporation of an existing targeting vector into ES cells.
Services (offered together or separately):
- Establishment of the complete molecular map of any type of BAC - derived or traditional gene targeting vector according to its basic design, (by the PGT Core).
- Targeting vector construction using the powerful method of BAC recombineering, or long range PCR (in collaboration with the DNA Sequencing Facility).
- Electroporation of gene targeting vectors into one of several C57BL6 or 129 mouse ES cell lines and selection with suitable drugs, depending on the vector (by the PGT Core).
- Full-service screening of the resulting ES cell clones by genomic Southern and genomic PCR (by the PGT Core).
- Karyotyping of correct recombinant ES cell clones (by the PGT Core).
- Expansion of ES clones for injection into blastocysts (in collaboration with the Transgenic and Chimeric Mouse Facility).
Current Research: The PGT Core and Laboratory has been funded by the NIH to develop gene targeting methods for human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC). Please inquire with Dr. Tobias Raabe if you are interested in a potential academic collaboration.
Tobias Raabe, Ph.D.