We employ this more classical method for the alleles that are not yet accessible directly in the embryo through genome editing technologies.
ES cells are electroporated with a targeting construct and colonies isolated. The clones are screened, amplified, quality controlled and converted to mice by microinjection into blastocysts. Chimera are obtained and mated for transmission of the genetic modification.
We combine classical methods with current advances, employing transient expression of CRISPR/Cas9 reagents to enhance targeting efficiency for challenging projects.
We generate classical targeting vectors or employ varied targeting constructs from external sources, e.g. EUCOMM/KOMP collection to BACs.
New alleles are extensively validated, combining classical methods (i.e. Southern blot) with new technologies (i.e. ddPCR).