Intermediate Vectors

 

 

Intermediate vectors are precursors of final vectors and lack the central targeting cassette. They are distributed to users who want to assemble their own targeting vectors with specific features that are not present in the EUCOMM final vectors.

 

The figure below illustrates the final steps leading from the intermediate vector to the final targeting vector. The intermediate vector contains the 5’ and 3’ homology arms for homologous recombination and the critical exon. One loxP site (red triangle) is also present. The intermediate vector still lacks the targeting cassette and a backbone with the negative selection cassette (DTA).

 

 

EuMMCR distributes intermediate vectors to interested users, who would like to introduce the targeting cassette of their liking into the final targeting vector, e.g. fluorescent reporters (GFP, mCherry, etc.), dominant alleles, suicide genes (DTA), or any other gene cassette for ectopic expression.

 

It is not possible to use intermediate vectors directly for the generation of mutant ES cell clones via electroporation, since they are lacking key features of the final targeting vectors.