Penn Gene Targeting Core and Laboratory
- targeting vector design and construction,
- electroporation of the targeting vector into ES cells,
- Southern/PCR genotyping and karyotyping of the resulting ES clones,
- and finally injection into blastocysts (usually together with the Transgenic and Chimeric Mouse Facility).
It also provides general genomic Southern/PCR services for genotyping of mouse tails as well as any cell type from which sufficient genomic DNA can be obtained (human ES cells, iPSC, mesenchymal stem cells and others).
In the past three years (2011-2013) 49 total mouse projects (of these ~15 in the C57BL6 background) were completed. Of these projects 46 went germ line and the remaining 3 did not go germ line because of definite or likely embryonic lethality.
Examples of publications describing targeted mice generated by the Penn Gene Targeting Core and Laboratory are listed here. Publications detailing the expertise of the Core regarding stem cells and regulation of gene expression are shown here.
For researchers of the University of Pennsylvania and affiliated institutions, prices are about one third to one half below those of comparable commercial services. In fact, even for outside academic researchers prices are still usually below those of the average commercial services. Although we have set prices for specific services, overall charges for a project are strongly dependent on the individual nature of the project and therefore its best to get a quote. We also give volumne discounts for some services.
For further details see here.
|Establishment of molecular map of BAC-derived targeting vectors that are tailor made for the desired mutation and based on a preexisting basic biological design.|
|Construction of gene targeting vectors either at the Penn Gene Targeting Core, or at the DNA Sequencing Facility or at outside companies, depending on the project. Even the most complex knockin vectors, such as vectors that insert drug-inducible and tissue-specific transgenes into the ROSA 26 locus, can be generated.|
|Electroporation of finished targeting vectors into one of several 129 or C57BL6 mouse ES cell lines and selection with suitable drugs, depending on the vector. Other cell types can be transfected as well (please inquire with Dr. Tobias Raabe for specifics). Our ES lines exhibit high rates of germline transmission (for details see here).|
|Genomic Southern and genomic PCR screening of any cell type, including targeted ES lines, and identification of the correct mutation. Please note that PGT accepts any cell type from which sufficient genomic DNA can be isolated for genomic Southern and PCR genotyping.|
|Karyotyping of recombinant ES clones and expansion for injection into mouse blastocysts. The Penn Gene Targeting Core coordinates timing of the expansion with the Transgenic and Chimeric Mouse Facility, which performs the injection into mouse blastocysts.|
|Injection into mouse blastocysts. Done at the Transgenic and Chimeric Mouse Facility, which performs the injection into mouse blastocysts and produces the chimera.|
- NIH-sponsored Knockout Mouse Project (KOMP)
- Penn Diabetes and Endocrinology Research Center (DERC)
- National Heart Lung and Blood Institute (NHLBI)
- Penn Institute for Regenerative Medicine (Penn IRM)
- Human Hematopoietic Stem Cell Center of Excellence (Children's Hospital of Philadelphia)
- Penn DNA Sequencing Facility
- Penn Transgenic and Chimeric Mouse Facility