Gene Targeting in Embryonic Stem Cells (ES cells)

Short Description
This service offers the transfection of ES cells with a targeting vector or transgenic vector (including BAC) to generate gene targeted or transgenic mouse strains. The cost for one round of transfections (2x107 cells) is CHF 5´000,- (+MWSt, non-USZ/UZH) which consist of consumables (3´000,-) and personnel (2´000,-). To improve the chances for a successful experiment we require that the construct and the targeting strategy has been presented and discussed during the Mouse Makers Club (every second Thursday 16:30 at room 44-J-05 on Irchel campus). We will expand up to 5 homologous recombinants for further characterization and use for chimera generation. Additional clones will be expanded on request and charged as 'expansion'. Due to the high variability in surviving recombinants which depends on the vector and target gene we can only guarantee a successful transfection by routinely checking transfection efficiency through inclusion of a transfection control. We offer targeting in C57BL/6, 129Ola, F1(C57BL/6x129SV), and BALB/c ES cells. Depending on the ES cell line between 30 - 60 % of recombinant clones give germline. We recommend that you aggregate three clones. However, please note that even then statistically each 8th experiment requires a fourth clone to be aggregated (assuming a 50% germline rate).

Prerequisites
- Approval of your targeting strategy through the mouse makers club
(proof of working Southern blot, proof of integrity of targeting vector)
- 100 µg of targeting vector (DNA), linearized, proof of linearization
(agarose gel with cut versus uncut, 200ng of each)
- a completely filled out request form

Targeting_Form.rtf (RTF, 351 KB)

Important
We normally try to process your request as soon as they arrive. In case of a backlog of requests, the date of DNA receipt and the number of requests submitted by individual groups will determine the sequence in which the projects are carried out.

In case of questions, please contact Thorsten Buch at buch@immunology.uzh.ch (044 635 37 09) before submitting a request.