Vector Design and Creation
iTL creates its targeting constructs using the high fidelity Red/ET recombineering method. We isolate genomic BACs from our internal library and use homologous recombination to produce the vectors. Compared to the traditional PCR based process, BAC recombineering is a more robust and faster technique less likely to introduce unwanted mutations.
The general targeting vector creation process includes analysis of gene structure, screening strategy design, probe design and validation, isolation and confirmation of genomic BAC clone, insertion and confirmation of homology arms and a selection cassette flanked by FRT and/or loxP sites and preparation of DNA for electroporation.
iTL Delivers:
- : targeted region replaced with a Neomycin selection cassette
- : targeted region flanked by lox-P sites
- : conventional or conditionally introduced engineered mutations
- : cDNAs and/or reporters are inserted to replace an endogenous gene or be fused to endogenous gene for coexpression
- : cDNA inserted into specific locus for targeted transgenic overexpression