Expansion of Gene Trap ES cells

What are Gene Trap ES cells?
Gene-trapped embryonic stem cells are a fast and easy way to obtain gene-deficient or hypomorphic alleles. The laboratories of the International Gene Trap Consortium have randomly mutated ES cells by insertion of retroviral vectors with a promoter-less resistance gene. Thousands of insertion ES cell clones were analysed by either genomic or cDNA sequencing and these sequences were made available via the gss database (pubmed). Gene trap ES cells give gene deficient, hypomorphic, or dominant negative mouse mutants, dependent on the gene, the type of vector used, and the location of the insertion within the gene.

To generate knockout mice through gene traps, identify three gene trap ES cell clones with insertions in the gene of your interest. Blast your cDNA in the nucleotide blast of pubmed versus the 'gss' database and type in as entrez query 'trap'. Some gene traps are placed with their exact genomic integration point (thus sequence flanking the inserted virus is deposited in the database). It may therefore be advisable to run in a second attempt the genomic locus against the database. For the highest likelyhood of obtaining a deficiency allele choose insertions in the 5´ part of the gene.
Order the ES cells from the respective service and have them send to the AGTF (make sure the bill goes to your address and not the AGTF).
It is advisable to join the Mouse Makers Club to design a strategy for confirming the integration into the appropriate locus once the cells have been expanded.

Short Description
This service offers the thawing and expansion of a gene trapped ES cell clone. We will freeze at least 8 vials of the clone. Three vials and three cell pellets will be given to the researcher. The cell pellets can be used for preparation of DNA for Southern blot analysis or PCR. This service costs 1´200 CHF (+MWSt, non-USZ/UZH) per clone.

Prerequisites
- you have already identified gene trap ES cells
- a completely filled out request form:

Gene_Trap_expansion_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 Thorsten.Buch@neuroimm.uzh.ch (044 635 37 09) before submitting a request.