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:

• Conventional targeting vector: targeted region replaced with a Neomycin selection cassette

• Conditional knockout vector: targeted region flanked by lox-P sites

• Knockin point mutation(s): conventional or conditionally introduced engineered mutations

• Knockin human or murine cDNAs or reporter genes: cDNAs and/or reporters are inserted to replace an endogenous gene or be fused to endogenous gene for coexpression

• Targeted transgenic:  cDNA inserted into specific locus for targeted transgenic overexpression

Find out more about our vector design technology.